Changes in plant mitochondria ultrastructure and respiration intensity in altered gravity.

Analysis of experimental results involving both animals and plants showed that cell mitochondria considerably changed regarding organelles shape, matrix density, number and size of cristae. However, oxidative-reductive enzymes did not lose their activity (2). Moreover, there are some data concerning an increase in respiration intensity of plants grown on a horizontal clinostat (3) and in seedlings being on board of Biosatellite-11 (4). Similar rearrangements in mitochondria are also observed during clinorotation (5). Taking into account that energy supply of cells during their adaptation to various factors considerably depends on the functional activity of these organelles along with ultrastructure the activity of some enzymes of energetic and oxidative metabolism as well as the level of cell respiration in altered gravity was studied.

Peculiarities of ultrastructure of Chlorella cells growing aboard the Bion-10 during 12 days.

The ultrastructure of Chlorella cells grown in darkness on a solid agar medium with organic additions aboard the Bion-10 biosatellite was studied. Certain differences in submicroscopic organization of organelles in the experimental cells were revealed compared to the Earth control. The changes are registered mainly in ultrastructure of energetic organelles--mitochondria and plastids of the experimental cells, in particular, an increase of mitochondria and their cristae size, as well as an increase of the total volume of mitochondrion per cell were established. The decrease of the starch amount in the plastid stroma and the electron density of the latter was also observed. In many experimental cells, the increase of condensed chromatin in the nuclei has been noted. Ultrastructural rearrangements in cells after laboratory experiment realized according to the thermogram registered aboard the Bion-10 were insignificant compared to the flight experiment. Data obtained are compared to results of space flight experiments carried out aboard the Bion-9 (polycomponent aquatic system) and the orbital station Mir (solid agar medium).

Peculiarities of the submicroscopic organization of Chlorella cells cultivated on a solid medium in microgravity.

The submicroscopic organization of Chlorella vulgaris cells (strain LARG-1) growing over 30 days on a solid agarized medium aboard the orbital station "Mir" was studied. A number of differences in the ultrastructure of cells of the experimental population compared to the control has been revealed. Thus, changes in the membrane system of plastids, in particular, appearance of numerous vesicles of different diameter and outgrowths of the plastids and their contact with the plasmalemma as well as a considerable decrease of reserve polysaccharide number in the plastids. Moreover, an increase in the size of mitochondria, their cristae and lipid drops in cytoplasm, the formation of more complicated configuration folding of plasmalemma and appearance of small-granular material of mean electron density in the periplasmic space of Chlorella cells grown during space flight, are demonstrated. Comparative cytological analysis has revealed general regularities of rearrangements of the submicroscopic organization in Chlorella cells cultivated on both solid and semiliquid agarized nutrient media.

Calcium gradient in plant cells with polarized growth in simulated microgravity.

Plant cells characterized by apical growth, for example, root hairs and apical cells of moss protonema, are a convenient model to address the problem of gravity response mechanisms including initiation of cell polarity. The fluorescent calcium probe, chlorotetracycline, allowed us to display the calcium distribution gradient in these cells. Irradiation by red light led to a sharp decrease in the Ca2+ ion activity in cells. During clinostatting in darkness the pattern of calcium influx and distribution changes inconsiderably as compared with control; in root hairs calcium is detected mainly in their apices and bases as in control. Addition of chlorpromazine to the medium probably increases the influx and accumulation of Ca2+ ions. Under data obtained confirm speculations on the Ca2+ ion functional role for the apical growth of plant cells and may suggest the participation of gravity in redistribution or activation of ion channels, calcium channels included, in the plasmalemma.

Plant cell plasma membrane structure and properties under clinostatting.

Structural-functional organization of plasma membrane of pea roots seedling was investigated by methods of chemiluminescence, fluorescence probes, chromatography and freeze-fracture studies under normal conditions and clinostatting. Phase character of lipid peroxidation intensity was fixed. The initial phase of this process is characterized by lipid peroxidation decreasing with its next induction. The primary changes depending on free-radical mechanisms of lipid peroxidation were excellently revealed by chemiluminescence. Plasmalemma microviscosity increased on the average of 15-20% under microgravity at the initial stages of its phenomenon. There were major changes of phosphatidilcholine and phosphatidilethanolamine contents. The total quantity of phospholipids remained rather stable. Changes of phosphatide acid concentration point to degradation and phospholipids biosynthesis. There were increases of unsaturated fatty acids mainly at the expense of linoleic and linolenic acids and also a decrease of saturated fatty acid content at the expense of palmitic and stearic acids. Unsaturation index of fatty acids increased as well. On the whole fatty acid composition was variable in comparison with phospholipids. Probably it is one of mechanisms of maintaining of microviscosity within definite limits. Considerable structural changes in organization of plasmalemma protein-lipid complex were not revealed by the freeze-fracture studies.

Ultrastructural and growth indices of Chlorella culture in multicomponent aquatic systems under space flight conditions.

Submicroscopic organization of Chlorella cells cultivated under space flight conditions in three-component aquatic system has been studied. Comparison of the experimental cells with that of the controls revealed certain rearrangements of cell organelles particularly, a reduction in the amount of reserve polysaccharides in chloroplasts, increase of cell vacuolization and mitochondrion volume, complication configuration of plasmalemma evaginations and invaginations, and also disturbances in the process of cytokinesis. More over an increase in the number of Chlorella cells infected by bacteria was shown in the experimental variant. No considerable differences were established in the growth characteristics of the experimental and control populations. A comparative cytological analysis revealed general regularities of organelles in Chlorella cells cultivated under space flight condition in the uni- and multicomponent systems.

[Isolation of a biologically active poly(A)+RNA from tissues enriched with polysaccharides]. / Poluchenie biologicheski aktivnoi poli(A)+RNK iz tkanei, obogashchennykh polisakharidami.

A special technique is developed for isolation of biologically functional poly(A)+RNA from higher plant cells to start with some procedures used to isolate RNA of the eucaryote cells. The technique includes cell disruption with a 4 M guanidine thiocyanate solution, hot phenol extraction of RNA following proteinase K digestion and methoxyethanol treatment to remove polysaccharides. Finally, poly(A)+RNA was purified by oligo(dT)-cellulose chromatography. Using this technique poly(A)+RNA preparations were obtained from French bean embryo axis, maize and sugar beet seedings, potato tubers and seedlings. It is shown that poly(A)+RNA isolated from the plant cells directs protein synthesis in the cell-free system. This RNA may be also used as a matrix for synthesis of cDNA in the reverse transcription reaction.

Prospects of autotrophic link functioning in biological life-support systems based on cell biology studies.

On the basis of using of modern methods of cytological analysis the cell growth peculiarities, reproduction and structure of plant cells of different level organisation (unicellular algae, mosses, angiosperms, cell cultures of higher factors--weightlessness, hypogravity, magnetic fields of various intensity, vibration and acceleration were cleared. It is shown that many discovered biological effects of space flight are connected with the degree of complexity of the object organisation and intensified with the increasing duration of influence.

Peculiarities of genital organ formation in Arabidopsis thaliana (L) Heynh. under spaceflight conditions.

An experiment was carried out ahoard the Salyut 6 research orbital station on Arabidopsis thaliana cultivations. The seeds were sprouted in the Svetoblok 1 device which provides for plant growth in the agar medium under sterile conditions and at 4000 lux illumination. The experimental plants, as well as the controls, reached approximately the same developmental stages: both flowered and began to bear fruit. A microscopic examination of the generative organs in the control and experimental plants shows that in normally formed (by appearance) flower buds and flowers of the experimental plants, as distinct from the controls, there were no fertile elements of the adroecium and gynoecium. Degeneration of the latter occurred at different stages of generative organ development. Possible reasons for this phenomenon in plants grown under weightless conditions are considered.

Biological effects of weightlessness and clinostatic conditions registered in cells of root meristem and cap of higher plants.

Research in cellular reproduction, differentiation and vital activity, i.e. processes underlying the development and functioning of organisms, plants included, is essential for solving fundamental and applied problems of space biology. Detailed anatomical analysis of roots of higher plants grown on board the Salyut 6 orbital research station show that under conditions of weightlessness for defined duration mitosis, cytokinesis and tissue differentiation in plant vegetative organs occur essentially normally. At the same time, certain rearrangements in the structural organization of cellular organelles--mainly the plastid apparatus, mitochondria, Golgi apparatus and nucleus--are established in the root meristem and cap of the experimental plants. This is evidence for considerable changes in cellular metabolism. The structural changes in the subcellular level arising under spaceflight conditions are partially absent in clinostat experiments designed to simulate weightlessness. Various clinostatic conditions have different influences on the cell structural and functional organization than does space flight. It is suggested that alterations of cellular metabolism under weightlessness and clinostatic conditions occur within existing genetic programs.

Biological studies of Chlorella pyrenoidosa (strain LARG-1) cultures grown under space flight conditions.

This paper reports data from an electron microscopic analysis of Chlorella pyrenoidosa (LARG-1) cultures after five days of growth in the dark on a semiliquid mineral/glucose medium in the IFS-2 device on board the Soyuz 27-Salyut 6-Soyuz 28 orbital research complex (Soviet-Czechoslovak experiment "Chlorella-1"). For space flight of five days duration the ultrastructural cellular organization of the flight and control cultures were similar. This testifies to normal cell function.

Growth and cell structure of Proteus vulgaris when cultivated in weightlessness in the Cytos apparatus.

Growth data and electron-microscopic analyses are presented for Proteus vulgaris cultures which were grown during space flight in polyethylene packets in a semisolid medium with Tryptose for 96 h. In the suboptimal culture conditions the growth and morphological characteristics of the flight and ground control variants were nearly identical, but we were able to detect a number of differences between the cellular ultrastructure of these variants. These differences testify to changes in the bacterial cell metabolism during space flight.

[Electron microscopic study of protoplasts of mesophyll tobacco. I. Ultrastructural and cytochemical study of tobacco protoplasts at early stages of cultivation]. / Elektronnomikroskopicheskoe izuchenie protoplastov mezofilla tabaka. Soobshchenie I. Ultrastrukturnoe i tsitokhimicheskoe issledovanie protoplastov tabaka na rannikh stadiiakh kul'tivirovaniia.

Ultrastructural changes in tobacco mesophyll protoplasts during the first three days of cultvation were studied. Localization of catalase and peroxidase activities in the freshly isolated protoplasts and 2 days after their cultivation was detected by the cytochemical 3,3'-diaminobenzidine method. A conclusion is drawn that the observed changes in the fine structure and localization of the enzymic activity are due to the following processes: reparation of disturbances caused by the isolation procedure, regeneration of the cell wall and rapid dedifferentiation of the initial parenchymatic leaf cells.