The accumulation of steviol glycosides (SGs) in cells of Stevia rebaudiana Bertoni both in vivo and in vitro was related to the extent of the development of the membrane system of chloroplasts and the content of photosynthetic pigments. Chloroplasts of the in vitro plants, unlike those of the intact plants, had poorly developed membrane system. The callus cells grown in the light contained proplastids of almost round shape and their thylakoid system was represented by short thylakoids sometimes forming a little number of grana consisting of 2 -3 thylakoids. In cells of the etiolated in vitro regenerants and the callus culture grown in the dark, only proplastids practically lacking the membrane system were observed. All the chloroplasts having developed thylakoids and forming at least a little number of grana were equipped with photochemically active reaction centers of photosystems 1 and 2. Leaves of in vivo plants accumulated greater amount of the pigments than leaves of the in vitro plants. In both the callus culture grown in the light and the etiolated in vitro regenerants, the content of the pigments was one order of magnitude lower than that in leaves of the intact plants. The callus tissue grown in the dark contained merely trace amounts of the pigments. Leaves of the intact and the in vitro plants did not exhibit any significant differences in photosynthetic O 2 evolution rate. However, photosynthetic O 2 evolution rate in the callus cells was much lower than that in the differentiated plant cells. The in vitro cell cultures containing merely proplastids did not practically produce SGs. However, after transferring these cultures in the light, both the formation of chloroplasts and the production of SGs in them were detected.Additional key words: photosynthetic rate, photosynthetic pigments, rebaudiosides A and C, stevioside.

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The epidermal salt glands of the leaf of Distichlis spicata 'Yensen 4a' (Poaceae) have a direct contact with one or two water-storing parenchyma cells, which act as collecting cells. A vacuole occupying almost the whole volume of the collecting cell has a direct exit into the extracellular space (apoplast) through the invaginations of the parietal layer of the cytoplasm, which is interrupted in some areas so that the vacuolar-apoplastic continuum is separated only by a single thin membrane, which looks as a valve. On the basis of ultrastructural morphological data (two shapes of the extracellular channels, narrow and extended, are found in basal cells), the hypothesis on the mechanical nature of the salt pump in the basal cell of Distichlis leaf salt gland is proposed. According to the hypothesis, a driving force giving ordered motion to salt solution from the vacuole of the collecting cell through the basal cell of the salt gland to cap cell arises from the impulses of a mechanical compression-expansion of plasma membrane, which penetrates the basal cell in the form of extracellular channels. The acts of compression-expansion of these extracellular channels can be realized by numerous microtubules present in the basal cell cytoplasm.

Effects of knockout of the gene encoding α carbonic anhydrase 4 (α CA4) on growth and photo synthesis of Thale cress (Arabidopsis thaliana (L.) Heynh., var. Columbia) were investigated. The shoot weight of mutant plants was found to be higher than in wild type plants, and the leaves of mutants were enriched in starch content. The electron microscopy study revealed a considerable increase in the number and size of starch grains in chloroplasts of mutant plants. Comparison of wild type and mutant plant leaves in terms of chlorophyll a fluorescence, coefficient of photochemical fluorescence quenching, effective quantum yield of photosystem II reaction, and nonphotochemical fluorescence quenching under steady state illumination and saturating CO 2 content in the air led to the proposal that α CA4 participates in the development of non photochemical energy dissipation by accelerating the supply of protons for activation of violaxanthin deep oxidase and for structural changes in the light harvesting complexes.

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The structural reorganization of pea thylakoid systems in response to osmotic shock in a wide range of temperatures (36-70 °C) was studied. At temperatures 40-46 °C, the configuration of thylakoid systems changed from a flattened to a nearly round, whereas thylakoids themselves remained compressed. The percentage of thylakoids stacked into grana at 44 °C decreased from 71 % in the control to 40 % in experimental samples, reaching 59 % at 48 °C. At 44 °C and above, thylakoid systems ceased to respond to the osmotic shock by disordering, in contrast to what happened at lower temperatures (36-43 °C) and in the control, and retained the configuration inherent in thylakoid systems at these temperatures. At 50 °C and above, the packing of thylakoids in grana systems changed, and thylakoids formed extended strands of pseudograna. Simultaneously, single thylakoids formed a network of anastomoses through local fusions. At temperatures of 60-70 °C, thylakoid systems appeared as spherical clusters of membrane vesicles with different degree of separation.Additional key words: osmotic shock; pea; Pisum; ultrastructure of thylakoid systems.

Specific temperature, storage times, and medium composition enable initiation of regular arrays of intramembranous particles on the exoplasmic fracture face during prolonged storage of isolated chloroplasts at 4 °C, producing about 2 – 10 regular arrays with 2 – 30 particles in each array, with a period of about 36 nm, oriented in 1 – 4 directions. The particle sizes do not change throughout the time of storage (1 – 4 weeks). The second type of particle regularity arises during prolonged storage of chloroplasts in greater than 1 M sucrose at −18 °C. Rounded areas of small particles tightly packed into paracrystalline arrays are found among less densely packed particles. The density of small particles is 4700 particles/μm2, and the mean size is 11 nm, whereas the particle density of the background is 1600 particles/μm2 with a mean particle size of 13 nm compared with 1200 particles/μm2 and mean size 16 nm in fresh chloroplasts. Based on the reduction of particle sizes and manner of packing on the fracture face, it is proposed that the small particles are a light-harvesting complex, separate from photosystem II and aggregated into paracrystalline arrays. The thylakoid lipids may participate in formation of particle regularity. Key words: thylakoid membrane, freeze fracture, particle regularity, low temperatures.

The comparative study of biochemical and ultrastructure features in senescing sugar beet (Beta vulgaris L.) leaves was carried out. One group of plants was grown under normal conditions in washed river sand and poured in turn with nitrate-containing mineral solution or water (N plants). Another group of plants, after 1 month of normal growth, was further grown with nitrate omitted in the nutritive solution (defN plants). The starting point of normal leaf senescence in N plants was identified by the maximal content of soluble protein. Soluble carbohydrate pools were statistically constant in senescing N plants, whereas glucose pools varied noticeably. A decrease in the contents of soluble protein and chlorophyll (a + b) in the course of senescing was typical for N plant leaves. The cell membrane in N plant leaves remained mostly intact; the central vacuoles in the leaf cells were large, and their membranes remained intact. The chloroplasts and mitochondria in senescing N plant leaves became swollen. The vesicles that were present in the cytoplasm of N plant leaves were especially large in the oldest leaves. It was concluded that senescing of sugar beet leaves at sufficient nitrate nutrition occurs according to a "vacuolar" scenario. In the case of nitrate deficiency, the content of soluble carbohydrates in defN leaves first reached maximum and then decreased in older leaves; the protein and chlorophyll (a + b) contents were totally lower than those in normal leaves and continuously decreased during the experiments. Chloroplasts in mesophyll cells of defN plant leaves became more rounded; starch grains in chloroplasts degraded and the number and size of lipid globules increased. The multitude of membrane impairments and lots of large vesicles-"crystals" appeared during the experiment. The results showed the controlling action of nitrogen nutrition in the senescing of sugar beet leaves.