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Biomedical subjects

E L Kordyum

Publications and source records attributed to E L Kordyum.

At least 19 recordsLinked to original sources

Application of GFP technique for cytoskeleton visualization onboard the International Space Station.

Cytoskeleton recently attracted wide attention of cell and molecular biologists due to its crucial role in gravity sensing and trunsduction. Most of cytoskeletal research is conducted by the means of immunohistochemical reactions, different modifications of which are beneficial for the ground-based experiments. But for the performance onboard the space vehicles, they represent quite complicated technique which requires time and special skills for astronauts. In addition, immunocytochemistry provides only static images of the cytoskeleton arrangement in fixed cells while its localization in living cells is needed for the better understanding of cytoskeletal function. In this connection, we propose a new approach for cytoskeletal visualization onboard the ISS, namely, application of green fluorescent protein (GFP) from Aequorea victoria, which has the unique properties as a marker for protein localization in vivo. The creation of chimerical protein-GFP gene constructs, obtaining the transformed plant cells possessed protein-GFP in their cytoskeletal composition will allow receiving a simple and efficient model for screening of the cytoskeleton functional status in microgravity.

Actin Cytoskeleton↗

Organization of cytoskeleton during differentiation of gravisensitive root sites under clinorotation.

Key role in cell gravisensing is attributed to the actin cytoskeleton which acts as a mediator in signaling reactions, including graviperception. Despite of increased attention to the actin cytoskeleton, major gaps in our understanding of its functioning in plant gravisensing still remain. To fill these gaps, we propose a novel approach focused on the investigation of actin involvement in the development of columella cells and cells in the transition zone of roots submitted to clinorotation. Both statocytes and cells in the transition zone represent the postmitotic cells which take origin in root meristems and are specified into graviperceptive (root cap) and gravireacting (transition zone) root tissues. The aim of the research was to investigate and compare the microfilament arrangements in root cap statocytes and peripheral root tissues (epidermis and cortex cells) in the transition zone and to find out how the actin cytoskeleton is involved in their specification under clinostat conditions. So far, our experiments have shown that under clinorotation the cytoplasmic microfilament network in the cortex cells in the transition zone is significantly enhanced. It is suggested that more abundant cytoplasmic microfilaments could strengthen the cortical actin cytoskeleton arranged parallel with the cortical microtubules, which are found to be partially disorganized in this area. Due to microtubule disorganization, the functioning of cellulose-synthesizing machinery and proper deposition of cell wall might be affected and could cause the alterations in the growth mode. But, in our case growth of the cells in the transition zone under clinorotation was rather stable. Due to our opinion, general stability of cell growth under clinorotation is promoted by mutual functional interrelation between actin and tubulin cytoskeletons. It is suggested that a strengthened cortical actin cytoskeleton restricts the cell growth instead of disorganized microtubules.

Actins↗

Alterations of the intranucleolar DNA localization caused by simulated microgravity.

The functions of the nucleolus responsible for the biosynthesis of ribosomes in altered gravity are still unclear. The location of nucleolar DNA both in control and simulated microgravity with fluorescent and immunogold cytochemistry was investigated. We determined firstly the redistribution of rDNA in the nucleolar components accompanied with the changes in their ultrastructure. According to it, lowering the level of rDNA transcription in simulated microgravity was firstly proposed.

Cell Nucleolus↗

Changes in vacuolation in the root apex cells of soybean seedlings in microgravity.

Changes in the vacuolation in root apex cells of soybean (Glycine max L. [Merr.]) seedlings grown in microgravity were investigated. Spaceflight and ground control seedlings were grown in the absence or presence of KMnO4 (to remove ethylene) for 6 days. After landing, in order to study of cell ultrastructure and subcellular free calcium ion distribution, seedling root apices were fixed in 2.5% (w/v) glutaraldehyde in 0.1 M cacodylate buffer and 2% (w/v) glutaraldehyde, 2.5% (w/v) formaldehyde, 2% (w/v) potassium antimonate K[Sb(OH)6] in 0.1 M K2HPO4 buffer with an osmolarity (calculated theoretically) of 0.45 and 1.26 osmol. The concentrations of ethylene in all spaceflight canisters were significantly higher than in the ground control canisters. Seedling growth was reduced in the spaceflight-exposed plants. Additionally, the spaceflight-exposed plants exhibited progressive vacuolation in the root apex cells, particularly in the columella cells, to a greater degree than the ground controls. Plasmolysis was observed in columella cells of spaceflight roots fixed in solutions with relatively high osmolarity (1.26 osmol). The appearance of plasmolysis permitted the evaluation of the water status of cells. The water potential of the spaceflight cells was higher than the surrounding fixative solution. A decrease in osmotic potential and/or an increase in turgor potential may have induced increases in cell water potential. However, the plasmolysed (i.e. non-turgid) cells implied that increases in water potential were accompanied with a decrease in osmotic potential. In such cells changes in vacuolation may have been involved to maintain turgor pressure or may have been a result of intensification of other vacuolar functions like digestion and storage.

Calcium↗

Calcium signaling in plant cells in altered gravity.

Changes in the intracellular Ca2+ concentration in altered gravity (microgravity and clinostating) evidence that Ca2+ signaling can play a fundamental role in biological effects of microgravity. Calcium as a second messenger is known to play a crucial role in stimulus-response coupling for many plant cellular signaling pathways. Its messenger functions are realized by transient changes in the cytosolic ion concentration induced by a variety of internal and external stimuli such as light, hormones, temperature, anoxia, salinity, and gravity. Although the first data on the changes in the calcium balance in plant cells under the influence of altered gravity have appeared in 80th, a review highlighting the performed research and the possible significance of such Ca2+ changes in the structural and metabolic rearrangements of plant cells in altered gravity is still lacking. In this paper, an attempt was made to summarize the available experimental results and to consider some hypotheses in this field of research. It is proposed to distinguish between cell gravisensing and cell graviperception; the former is related to cell structure and metabolism stability in the gravitational field and their changes in microgravity (cells not specialized to gravity perception), the latter is related to active use of a gravitational stimulus by cells presumebly specialized to gravity perception for realization of normal space orientation, growth, and vital activity (gravitropism, gravitaxis) in plants. The main experimental data concerning both redistribution of free Ca2+ ions in plant cell organelles and the cell wall, and an increase in the intracellular Ca2+ concentration under the influence of altered gravity are presented. Based on the gravitational decompensation hypothesis, the consequence of events occurring in gravisensing cells not specialized to gravity perception under altered gravity are considered in the following order: changes in the cytoplasmic membrane surface tension --> alterations in the physicochemical properties of the membrane --> changes in membrane permeability, ion transport, membrane-bound enzyme activity, etc. --> metabolism rearrangements --> physiological responses. An analysis of data available on biological effects of altered gravity at the cellular level allows one to conclude that microgravity environment appears to affect cytoskeleton, carbohydrate and lipid metabolism, cell wall biogenesis via changes in enzyme activity and protein expression, with involvement of regulatory Ca2+ messenger system. Changes in Ca2+ influx/efflux and possible pathways of Ca2+ signaling in plant cell biochemical regulation in altered gravity are discussed.

Calcium↗

Effect of simulated microgravity on potato minituber formation and structure.

The effect of long-term clinorotation on potato minituber formation and the structural-functional organization of storage parenchyma cell in minitubers has been studied by using methods of organ culture in vitro, light- and electron microscopy, biochemistry as well as phenological observation. It was established some acceleration of growth, changes in the parenchyma cell ultrastructure and in the starch content as well as an intensification of phosphorylase activity in the storage tissue of minitubers under the influence of simulated microgravity.

Journal Article↗

The interaction of microgravity and ethylene on the ultrastructure cell and Ca2+ localization in soybean hook hypocotyl.

Calcium ions are secondary messenger in numerous cellular processes of plant grown at 1 g. Ca2+ are connected with oxygen atoms, of pectin carboxy groups and/or with H(+)-groups of protein (Roux and Slocum, 1982; Hepler and Wayne, 1985). The influence of altered gravity on the calcium balance in some cells is established. The increased synthesis of ethylene in plant grown in microgravity caused the change of the structural-functional organization of cell (Hensel and Iversen, 1980; Hilaire et al., 1996). Available data put the new question: how do high ethylene level and microgravity influence on the redistribution of Ca2+ in cell of seedling in early stage of growth? Therefore, the goal of our data was the comparable study of the cell ulltrastructure and localization of Ca2+ in hook hypocotyl of soybean seedling under interaction of microgravity and ethylene.

Calcium↗

Microgravity mediated changes in phytoferritin accumulation in soybean root cap cells.

Phytoferritin is an iron-protein complex analogous to the ferritin found in mammalian, bacteria and fungi cells. Phytoferritin molecules are large proteins, about 10.5 nm in diameter, visualised in an electron microscope as discrete, electron dense particles with iron-containing core, where several thousand atoms of iron lie within the proteinaceous shell (apoferritin). In higher plants, a plastid stroma is the site of phytoferritin storage. Phytoferritin is seen in all types of plastids. It is considered to be a mechanism used by cells to store iron in a non-toxic form. Phytoferritin-bound iron may subsequently be used to form iron-containing components. It was shown that low levels of phytoferritin are synthesised in normal green leaves, whereas chlorotic leaves do not have a measurable amount of phytoferritin and leaves of iron-loaded seedlings contain a high level of total iron, and phytoferritin well-filled by iron. Phytoferritin accumulation was observed in photosynthetic inactivity chloroplasts during senescence and disease. In this study we analised the effects of microgravity and ethylene on production of phytoferritin in the root cap columella cells of soybean seedlings.

Ethylenes↗

Gravitropism and phototropism in protonemata of the moss Pohlia nutans (Hedw.) Lindb.

The gravitropism of protonemata of Pohlia nutans is described and compared with that of other mosses. In darkness, protonemata showed negative gravitropism. Under uniform illumination they grew radially over the substrate surface, whereas unilateral illumination induced positive phototropic growth. Gravitropism was coupled with starch synthesis and amyloplast formation. Protonematal gravitropic growth is more variable than the strict negative gravitropism of Ceratodon chloronema.

Bryopsida↗

High temperature alters the growth reaction of Pottia protonemata.

Under gravistimulation, dark-grown protonemata of Pottia intermedia revealed negative gravitropism with a growth rate of approximately 28 micrometers hour-1 at room temperature (20 degrees C). In 7 days, the protonema formed a bundle of vertically oriented filaments. At an elevated temperature (30 degrees C), bundles of vertically growing filaments were also formed. However, both filament growth rate and amplitude of the gravicurvature were reduced. Red light (RL) irradiation induced a positive phototropism of most apical protonemal cells at 20 degrees C. In a following period of darkness, approximately two-thirds of such cells began to grow upward again, recovering their negative gravitropism. RL irradiation at the elevated temperature caused a partial increase in the number of protonemal cells with negative phototropism, but the protonemata did not exhibit negative gravitropism after transfer to darkness. The negative gravitropic reaction was renewed only when protonemata were placed at 20 degrees C. A dramatic decrease in starch amount in protonemal apical cells, which are sensitive to both gravity and light, occurred at the higher temperature. Such a decrease may be one of the reasons for the inhibition of the protonemal gravireaction at the higher temperature. The observation has a bearing on the starch-statolith theory.

Bryopsida↗

The gravireaction of Ceratodon protonemata treated with gibberellic acid.

Moss protonemata exhibit negative gravitropism and the amyloplasts of the apical cell seem to play a key role in protonemal gravisensitivity. However, the mechanisms of this process are still poorly understood. Previously, we have shown that Ceratodon protonemata grown on agar-medium demonstrated greater gravicurvature than protonemata grown on medium with 11 mM glucose. In this study, we have examined whether gibberellic acid (GA), which promotes alpha-amylase expression, influences graviresponse of C. purpureus protonemata (strains WT-4 and WT-U) and how this event interacts with exogenous soluble sugars. After gravistimulation the WT-4 strain curved about twice as fast as the WT-U strain. However, responses of both strains to added substances were similar. High concentration of glucose (0.11 M) caused a decrease in protonema curvature, while the same concentration of sucrose did not significantly change the angles of curvature compared with controls. GA at 0.1 mM and higher concentrations inhibited gravitropism, and caused some apical cells to swell. The possible involvement of the carbohydrates in gravitropism is discussed.

Bryopsida↗

DNA content and differentiation of root apical cells of Brassica rapa plants grown in microgravity.

Root cap is proposed to be a graviperceptive tissue in the plant root, and it is composed of several cell types. One such cell type, the columella cells, are thought to initiate the gravity-induced signal transduction cascade, and these cells arise from the activity of the meristematic zone of the root cap. There is, in fact, a continuum of cells in the central column of the root cap representing the meristematic cells, developing columella cells, mature cells, and those that will soon be sloughed off into the soil. In order to study the functional roles of the root cap cells in gravity-sensing, we compared the ultrastructural organization, differentiation, and DNA content in the meristematic, elongating, and differentiating cells of root tips in Brassica rapa plants grown in space microgravity and at 1g. The experiments were also designed to determine the reactions of root cap cells in both main roots (in which the original root cap was present in an embryonic form within the seed) and lateral roots (in which the root cap formed completely in space after seed germination on orbit) to the space microgravity. This study (ROOTS) was performed in collaboration with the B-PAC experiment on the Space shuttle "Columbia" mission STS-87 (Collaborative US/Ukrainian Experiment (CUE) during November 19-December 5, 1997.

Brassica↗

Some characteristics of photosynthetic apparatus under conditions of spaceflight.

During colonization of space by humans, the bioregenerative life-support systems on board the space ships will require the plants with a highly efficient photosynthesis, a process producing food and O2 and removing CO2-Therefore, in recent years the scientists increasingly focus the their attention to study on photosynthetic apparatus of plants grown in space. Although the available data are quite scanty and, at times, controversial, it is Considered that the space grown plants differ from around control plants by growth and development, many structural and functional indices and metabolism. Data exist showing changes in the chlorophyll (Chl) content, structure and number of chloroplasts in the cell, swelling of thylakoids and decrease in the number and size of starch grains in the chloroplasts (for reviews, see Halstead and Dutcher, 1987; Kordyum, 1997). The decrease of shoot fresh weight and reduction of CO2-saturated photosynthetic rate at saturating light intensities in space grown wheat plants in comparison with ground control have been reported by Tripathy et al. (1996). The thylakoids isolated from space grown plants displayed lower rates of electron transport through photosystems I and II (PSI and PSII) and in a whole chain. This study aimed to examine the electron transport rates through PSI and PSII in the isolated thylakoids, to elucidate if there are any differences in accumulation of thylakoid membranes between space grown Brassica rapa plants and ground control plants (based on Chl quantity) and to measure the Chl a/b ratio in isolated thylakoids. These studies were part of the Collaborative US/Ukrainian program during the STS-87 mission (1997).

Brassica↗

Plant reproduction systems in microgravity: experimental data and hypotheses.

Elucidation of the possibilities for higher plants to realize complete ontogenesis, from seed to seed, and to propagate by seeds in microgravity, is a fundamental task of space biology connected with the working of the CELSS program. At present, there are results of only 6 spaceflight experiments with Arabidopsis thaliana, an ephemeral plant which will flower and fruit in orbit. Morphogenesis of generative organs occurs normally in microgravity, but unlike the ground control, buds and flowers mainly contain sterile elements of the androecium and gynoecium which degenerate at different stages of development in microgravity. Cytological peculiarities of male and female sterility in microgravity are similar to those occurring naturally during sexual differentiation. Many of the seed formed in microgravity do not contain embryos. Hypotheses to explain abnormal reproductive development in microgravity are: 1) nutritional deficiency, 2) insufficient light, 3) intensification of the influence of the above-mentioned factors by microgravity, 4) disturbances of a hormonal nature, and 5) the absence of pollination and fertilization. Possible ways for testing these hypotheses and obtaining viable seeds in microgravity are discussed.

Ecological Systems, Closed↗

Gravi- and photostimuli in moss protonema growth movements.

Moss protonemal growth direction is controlled by at least three factors, photo-, gravi- and autotropism. It is possible to experimentally separate these factors and to control selectively their morphological appearance. In darkness protonema grow negatively gravitropically, and unilateral illumination initiated positive phototropism. Red light suppressed auto- and gravitropism, blue light suppressed only gravitropism. Green light allowed both gravi- and autotropism. The effect of light on gravitropism might involve changes in starch synthesis.

Bryopsida↗

Effects of microgravity on the susceptibility of soybean to Phytophthora sojae.

The study of pathogenicity of higher plants under conditions of microgravity is of great importance for the future production of food in space. Previous work suggests that microgravity affects both microbes and plants. Bacterial numbers increased after 17 days in an algae-bacterium association on the biosatellite "Kosmos-1887". This was speculated to result from an increase in the multiplication rate of the bacteria. Sporangia of both Actinomices brevis, in the shuttles "Soyuz-19" and "Appolon", and Phycomyces blakes, in biosatellite "Kosmos-936", formed after 10 days in microgravity. Sporangia did not form in the ground controls in the same time suggesting that the rate of fungal development is enhanced in microgravity. Plant responses to pathogens in microgravity have not been studied, however, microgravity profoundly impacts plant cell development, cytology, and physiology. In microgravity, developing cell walls are thinner and contain less lignin than ground-grown plants. The demonstrated effects of microgravity on both plants and microbes lead us to hypothesize that plants may be more susceptible to pathogens under conditions of microgravity. The aim of this study was to determine the influence of microgravity on the susceptibility of soybean to the fungal root rot pathogen, Phytophthora sojae.

Cytoplasm↗

Growth of Pottia intermedia protonemata in altered gravity.

Plants are immobile; therefore, they are oriented in space due to growth movements--tropisms. The latter occur in response to environmental stimuli such as gravity (gravitropism), light (phototropism), chemical compounds or water (chemo- and hydrotropisms). Gravity is the only force that was impossible to control. The moss protonemata are among the limited group of plant objects with tip growth. What is unique about this structure is that protonemal apical cells both sense and respond to gravity. It is considered that the apical cell perceives gravity through amyloplasts (Sack, 1993; Chaban, 1996). Although the dynamics of protonemata negative gravitropism in different moss species was studied in detail, the role of gravity in both the structural polarity of apical cells and the formation of protonematal mat with circular symmetry is completely unexplored. Using the unique possibility to fly the moss on the space shuttle (STS-87) we aimed in this study to analyze the character of the interaction of gravity with light and endogenous factors in the pattern of protonemata space orientation.

Bryopsida↗