Emilia L. Apostolova

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.

Electric properties of thylakoid membranes from pea mutants with modified carotenoid and chlorophyll-protein complex composition

Surface electric properties of thylakoid membranes from wild type and two mutant forms, Coeruleovireus 2/16 and Costata 2/133, of pea are investigated by electric light scattering and microelectrophoretic measurements. Characterization of the chlorophyll–protein complexes in thylakoid membranes reveals that the relative ratio of oligomeric (LHC II1) to monomeric (LHC II3) forms of the light-harvesting Chl a/b complex of Photosystem II is lower (3.34) in 2/133 mutant and higher (6.62) in 2/16 mutant than in wild type (4.57). This is accompanied by elevated amounts and a considerable reduction of all carotenoids in 2/16 and 2/133 mutant, respectively, as compared to the wild type. The concomitant variations of the permanent dipole moment (transversal charge asymmetry), electric polarizability and electrokinetic charge of the thylakoid membranes from both the mutants are discussed in terms of the differences in the supramolecular (oligomeric) organization of the light-harvesting complexes II within the photosynthetic apparatus.

Factors influencing PS II particle array formation in Arabidopsis thaliana chloroplasts and the relationship of such arrays to the thermostability of PS II

Chloroplasts of the thermal-stable fatty acid desaturase mutants JB67 and LK3 of Arabidopsis thaliana are characterised by the presence of regular arrays of freeze-fracture particles associated with the core and light-harvesting antennae of Photosystem II (Tsvetkova et al. (1994) Biochim. Biophys. Acta 1192, 263–271). Similar arrays were found to be induced in the membranes of chloroplasts isolated from wild-type plants by resuspending the chloroplasts in media containing Tricine and/or high concentrations of compatible co-solutes such as sorbitol. The thermal stability of their chloroplasts was also increased under such conditions. The increased tendency to form PS II particle arrays, and the enhanced thermal stability of PS II, in chloroplasts isolated from the mutants and wild-type chloroplasts suspended in different media, appear to be reflections of the increased stability of protein-protein interactions between and within PS II units, respectively. The role of lipids in determining the formation of freeze-fracture particle arrays in the mutants is discussed in terms of the observed changes in lipid composition and their possible role in the control of lipid/protein synthesis.

Relationship between the degree of carotenoid depletion and function of the photosynthetic apparatus

Fluridone, an inhibitor of the carotenoid biosynthesis, was used to study the relationship between the degree of carotenoid depletion and the function of the photosynthetic apparatus. The data reveal that, at a small reduction of the carotenoid content (25% decrease of the total carotenoids), the PSII and PSI (oxidation of P700 by far-red light) photochemistry is not influenced, while the oxygen evolution is strongly inhibited. Further reduction of the total carotenoid content (more than 40%) leads to decrease of the chlorophyll content and inhibition of the functions of both photosystems as the effect on the photosynthetic oxygen evolution and primary photochemistry is stronger than the effect on P700 oxidation. The analysis of the oxygen production under continuous illumination and flash oxygen yields suggests that the inhibition of the oxygen evolution is caused mainly by the damage of PSIIalpha centers.

Action and target sites of nitric oxide in chloroplasts

Nitric oxide (NO) is an important signalling molecule in plants under physiological and stress conditions. Here we review the influence of NO on chloroplasts which can be directly induced by interaction with the photosynthetic apparatus by influencing photophosphorylation, electron transport activity and oxido-reduction state of the Mn clusters of the oxygen-evolving complex or by changes in gene expression. The influence of NO-induced changes in the photosynthetic apparatus on its functions and sensitivity to stress factors are discussed.

Effects of exogenous 24-epibrassinolide on the photosynthetic membranes under non-stress conditions

In the present work the effects of exogenous 24-epibrassinolide (EBR) on functional and structural characteristics of the thylakoid membranes under non-stress conditions were evaluated 48 h after spraying of pea plants with different concentrations of EBR (0.01, 0.1 and 1.0 mg.L(-1)). The results show that the application of 0.1 mg.L(-1) EBR has the most pronounced effect on the studied characteristics of the photosynthetic membranes. The observed changes in 540 nm light scattering and in the calorimetric transitions suggest alterations in the structural organization of the thylakoid membranes after EBR treatment, which in turn influence the kinetics of oxygen evolution, accelerate the electron transport rate, increase the effective quantum yield of photosystem II and the photochemical quenching. The EBR-induced changes in the photosynthetic membranes are most probably involved in the stress tolerance of plants.

Photoelectron transport ability of chloroplast thylakoid membranes treated with NO donor SNP: Changes in flash oxygen evolution and chlorophyll fluorescence

The nitric oxide (NO) donor sodium nitroprusside (SNP) is frequently used in plant science in vivo. The present in vitro study reveals its effects on the photosynthetic oxygen evolution and the chlorophyll fluorescence directly on isolated pea thylakoid membranes. It was found that even at very low amounts of SNP (chlorophyll/SNP molar ratio∼67:1), the SNP-donated NO stimulates with more than 50% the overall photosystem II electron transport rate and diminishes the evolution of molecular oxygen. It was also found that the target site for SNP-donated NO is the donor side of photosystem II. Compared with other NO-donors used in plant science, SNP seems to be the only one exhibiting stimulation of electron transport through photosystem II.

Effect of phosphatidylglycerol depletion on the surface electric properties and the fluorescence emission of thylakoid membranes

To explore the possible effect of phosphatidylglycerol (PG) on the surface electric properties and chlorophyll fluorescence characteristics we used electric light scattering technique and 77K chlorophyll fluorescence of thylakoid membranes from a cyanobacterium, Synechocystis PCC6803 (wild type) and its pgsA mutant defective in PG synthesis. We found a strong decrease in the permanent and induced electric dipole moments of the mutant thylakoids, following long-term PG depletion parallel with a decrease of the emission peak from PSI and an increase of the emission peak from PSII. Partial recovery of the electric state of thylakoid membranes was observed at re-addition of PG to the mutant cells depleted of PG for 21 days. This change in the electric dipole moments is probably due to a decrease in PG content and progressive structural alterations in the macroorganization of the photosynthetic complexes induced by PG deprivation. Our results suggest that the depletion of a lipid, which carries a negative charge, despite its small contribution to the overall lipid content, significantly perturbs the surface charge of the membranes. These changes are related with the chlorophyll fluorescence emission ratios of two photosystems and may partly explain our earlier results concerning the PG requirement for the function and assembly of photosystems I and II reaction centers.

Effects of salinity on the photosynthetic apparatus of two Paulownia lines

The effects of soil salinity on the functional activity of photosynthetic apparatus and pigment composition of two Paulownia lines (Paulownia tomentosa x fortunei and Paulownia elongata x elongata) were investigated. PAM chlorophyll fluorescence measurements revealed that salinity leads to: (i) an increase of the photochemical quenching coefficient (qP) and the linear electron transport rate (ETR) in both lines of Paulownia, while the maximum quantum yield of the primary photochemistry of PSII in the dark adapted state (Fv/Fm) was unaffected; (ii) improved the efficiency of the photochemical energy conversion (ФPSII); (iii) an impact on the chlorophyll fluorescence decrease ratio (RFd), which correlates to the net CO2 assimilation rate; (iv) an impact on [Formula: see text] reoxidation. The analysis of the kinetics of P700(+) reduction upon turning off far-red irradiation revealed that salinization lead to a delay of the cyclic electron transport around PSI in both studied lines as the effect on this process is more pronounced in P. tomentosa x fortunei than in (in comparison with) P. elongata x elongata. The present experimental results suggested high salt tolerance of the studied lines Paulownia, but P. tomentosa x fortunei is more tolerant to salinity than P. elongata x elongata. Molecular mechanisms involved in the Paulownia response to the soil salinity are discussed.

SURFACE ELECTRIC PROPERTIES OF THYLAKOID MEMBRANES FROM ARABIDOPSIS THALIANA MUTANTS

Electric light scattering measurements of thylakoid membranes from wild type and two mutant forms (JB67 and LK3) of Arabidopsis thaliana have shown that application of external electric pulses induces electric dipole moments of different origin. The asymmetric surface charge distribution and electric polarizability are significantly altered by the lipid modification. Mild trypsin treatment of Arabidopsis thylakoids leading to digestion of small polypeptides from the light-harvesting chlorophyll a/b protein complex of photosystem II (LHCP II) gives evidence for a lower content of LHCP II in the mutant forms. The results demonstrate the significance of the level of thylakoid lipid unsaturation in determining the surface charge distribution through changes either in the pigment-protein content and membrane appression induced by the lipid modification or in the exposure of charged polypeptides on the thylakoid membrane surface(s) arising from alteration of the lipid geometry.