Katerina Stoitchkova

Changes in the energy distribution between chlorophyll-protein complexes of thylakoid membranes from pea mutants with modified pigment content. I. Changes due to the modified pigment content.

The low-temperature (77 K) emission and excitation chlorophyll fluorescence spectra in thylakoid membranes isolated from pea mutants were investigated. The mutants have modified pigment content, structural organization, different surface electric properties and functions [Dobrikova et al., Photosynth. Res. 65 (2000) 165]. The emission spectra of thylakoid membranes were decomposed into bands belonging to the main pigment protein complexes. By an integration of the areas under them, the changes in the energy distribution between the two photosystems as well as within each one of them were estimated. It was shown that the excitation energy flow to the light harvesting, core antenna and RC complexes of photosystem II increases with the total amount of pigments in the mutants, relative to the that to photosystem I complexes. A reduction of the fluorescence ratio between aggregated trimers of LHC II and its trimeric and monomeric forms with the increase of the pigment content (chlorophyll a, chlorophyll b, and lutein) was observed. This implies that the closer packing in the complexes with a higher extent of aggregation regulates the energy distribution to the PS II core antenna and reaction centers complexes. Based on the reduced energy flow to PS II, i.e., the relative increased energy flow to PS I, we hypothesize that aggregation of LHC II switches the energy flow toward LHC I. These results suggest an additive regulatory mechanism, which redistributes the excitation energy between the two photosystems and operates at non-excess light intensities but at reduced pigment content.

Selective Photobleaching of Chlorophylls and Carotenoids in Photosystem I Particles under High-Light Treatment

Photosystem I particles (PSI‐200) isolated from spinach leaves were studied by means of absorbance, 77K fluorescence and resonance Raman (RR) spectroscopy. The aim was to obtain better insight into the changes of the pigment spectral properties in those particles during prolonged exposure to high‐light intensities and to reveal the involvement of these pigments in the photoprotection of the PSI. During prolonged exposure to high‐light intensities of spinach PSI particles, a loss of a significant amount of photosynthetic pigments was observed. It was shown that various pigments exhibited different susceptibility to photodamage. In addition to bleaching of chlorophyll a (Chl a), bleaching of carotenoids was also clearly observed. RR technique allowed us to recognize the type and conformation of photobleached carotenoid molecules. Raman data revealed a nearly full photobleaching of the long‐wavelength lutein molecules. The observed similar bleaching rate of the lutein molecules and the most‐red shifted long‐wavelength Chl a, located in the antenna membrane protein Lhca4, suggested that these molecules are located closely. Our results showed that the photobleached antenna pigments and especially luteins and the most long‐wavelength absorbing chlorophylls are involved in photoprotection of PSI core complex.

Photoinduced changes in photosystem II pigments

The photosynthetic apparatus in higher plants performs two seemingly opposing tasks: efficient harvest of sunlight, but also rapid and harmless dissipation of excess light energy as heat to avoid deleterious photodamage. In order to study this process in pigment-protein supercomplexes of photosystem II (PSII), 77 K fluorescence and room temperature resonance Raman (RR) spectroscopy were applied to investigate the changes in structure and spectral properties of the pigments in spinach PSII membranes. The high-light treatment results in a strong quenching of the fluorescence (being largest when the excitation is absorbed by carotenoids) and a red-shift of the main maximum. Decomposition of the fluorescence spectra into four bands revealed intensive quenching of F685 and F695 bands, possible bleaching of chlorophyll a, enhanced extent of light harvesting complexes (LHCII) aggregation and increased energy transfer to aggregated LHCII. The analysis of RR spectra revealed the predominant contribution of s-carotene (s-Car) upon 457.8 and 488 nm excitations and lutein (Lut) at 514.5 nm. During prolonged exposure to strong light no significant bleaching of s-Car and weak photobleaching of Lut is observed. The results will contribute to the efforts to produce more efficient and robust solar cells when exposed to fluctuations in light intensity.

Light induced changes in Raman scattering of carotenoid molecules in Photosystem I particles

The photosynthetic antenna systems are able to regulate the light energy harvesting under different light conditions by dynamic changes in their protein structure protecting the reaction center complexes. The changes modulate the electronic structure of the main antenna pigments (chlorophylls and carotenoids) and distort the characteristic planar structure of carotenoids, allowing their forbidden out of plane vibrations. Electronic absorption and low-temperature resonance Raman spectroscopy were used to study the changes in composition and spectral properties of the major carotenoids in spinach Photosystem I particles due to high light treatment. The duration of the applied intensity of the white light (1800 &mgr;E m-2 s-1) was 30, 60 and 120 minutes. We used Raman scattering in an attempt to recognize the type and conformation of photobleached carotenoid molecules. The resonance Raman spectra were measured at 488 and 514.5 nm, coinciding with the absorption maximum positions of the carotenoids neoxanthin and lutein, correspondingly. The results revealed nearly a full photobleaching of the long wavelength lutein molecules, whereas the bleaching of neoxantin molecules is negligible. The involvement of these changes in the photoprotection and photoinactivation of the Photosystem I particles was discussed.

Relation Between Changes in Pigments’ Spectral Properties and Structural Distortions of Pigment Protein Complexes (abstract)

Scientists continue to investigate photosynthesis—nature’s process to efficiently regulate and store energy. To explore the mechanisms of regulation we used fluorescence, resonance Raman spectroscopy, and biochemical preparative methods. For detailed analysis we applied decomposition of the low temperature steady‐state fluorescence spectra. This allowed us to estimate the emission of distinct pigment protein complexes (PPC) and evaluate the fluorescence of various aggregation forms of the main light harvesting complex (LHCII), which plays a major role in the studied mechanisms. Resonance Raman spectroscopy revealed with precision the relation between changes in pigments’ spectral properties and structural distortions of PPC. It was shown that aggregation of LHCII led to out‐of‐plane distortion, not only of neoxanthin, but also of lutein molecules. This was enhanced when the complex was embedded in thylakoid membranes (TM). It was suggested that lutein molecules are more closely related to the process of a...