Distribution of Calcium Ions in Cells of the Root Distal Elongation Zone Under Clinorotation

Currently, calcium ions are known to play a crucial role in the vital activity of plant cells and in stimulus–response coupling for many environmental signals, altered gravity included. The available data on changes in Ca^2?+? distribution and concentration in the cells of different organisms influenced by altered gravity allow to suggest that microgravity affects the calcium messenger system, and provide new insight for the understanding of calcium-and gravity-dependent cellular processes. We have studied with confocal microscopy the distribution and relative content of calcium ions in the Beta vulgaris root distal elongation zone cells grown under slow horizontal clinorotation, reproducing one of the microgravity particularities, namely the absence of an orienting action of the gravity vector, compared to control conditions. We demonstrate that Ca^2?+? relative content is 1.3 times higher in the roots of seedlings grown upwards and 1.2 times higher in the seedlings grown downwards compared to the control. Based on obtained data, taking into account the specific physiological properties of cells in the distal elongation zone, it is supposed that, under clinorotation, enhanced Ca^2?+? relative content affects Ca^2?+?-dependent cytoskeleton reorganization involved in cell gravisensing in altered gravity.     

Clinorotation Effect on Response of Cress Leaves to Red and Far-Red Light

Responses of cress (Lepidium sativum L.) seedling leaves to separate and simultaneous illumination with red (660 nm) and far-red (735 nm) light were studied under fast clinorotation (50 rpm) and usual gravity (1g) conditions. The monochromatic light emitting diodes (LEDs) have been used for illumination of seedlings from above. The growth and spatial orientation of leaves and the location of presumable gravisensors in petioles were analysed. Clinorotation in the dark promoted the radial expansion of leaf lamina and unfolding of leaves. It was shown that clinorotation in red light inhibited significantly the elongation of petioles as compared with that under the action of gravity force. Simultaneous red and far-red illumination promoted the growth of petioles under clinorotation, but did not affect the orientation of laminas, which remained the same as of the 1-g control ones. Red light, applied separately and simultaneously with far-red light, guided the bending of laminas as well as the unfolding of leaf petioles in both usual and clinorotation conditions. Histological and cytological analyses of petioles revealed the presence of movable amyloplasts in endodermic cells in proximal region of petioles. Comparison of intracellular positioning of amyloplasts in petioles of leaves grown under clinorotation and the action of gravity allows a presumption that these plastids may be identified as gravisensors of garden cress leaves. The obtained data imply that clinorotation and exposition to monochromatic red light or simultaneous illumination by red and far-red light affect the elongation of petioles of cress seedling leaves. Spectral components guide the unfolding of laminas in a gravity-independent manner.     

Chlorophyll,Carotenoid and Anthocyanin Accumulation in Mung Bean Seedling Under Clinorotation

The accumulation of plant pigments in mung bean (Vigna radiata L.) seedlings was measured after clinorotation (2 rpm for 2-4 days), and compared to a stationary control. The pigments measured included chlorophyll and carotenoid in primary leaves, and the anthocyanin in seedlings. While significant changes in chlorophyll and carotenoid accumulation were not observed during the initial 2 to 4 days of cultivation, by day 4 the seedlings grown on the clinostat had lower levels of anthocyanin, compared to those in the control seedlings. To further detail the cause for the observed reduction in anthocyanin accumulation under altered gravity conditions, seedlings were grown in the presence of silver nitrate, a known ethylene inhibitor, for 4 days, since it is known ethylene has a negative impact on anthocyanin accumulation. Silver nitrate promoted anthocyanin accumulation in the clinostat seedlings, and as a result there was no significant difference between the control and clinostat seedlings in anthocyanin accumulation. The results suggest that slow clinorotation negatively impacts anthocyanin pigmentation in mung bean seedlings, with endogenous ethylene suspected to be involved in this.     

Comparison Study of Gravity-Dependent Displacement of Amyloplasts in Statocytes of Cress Roots and Hypocotyls

In this study, the kinetics of gravity-dependent movement of amyloplasts (statoliths) along root statocytes and hypocotyls (endodermis cells) has been analyzed and compared in order to testify cytoskeleton involvement in the displacement of statoliths in cress (Lepidium sativum L.) seedling statocytes. After 32 h of growth at 1 g or under a fast clinorotation (50 rpm), the seedlings were treated for 24 min as follows: exposition to clinorotation or 180° inversion and the action of gravitational force in root tip or hypocotyl tip direction. Statolith displacement was studied by light microscopy on semi-thin longitudinal sections of hypocotyls and root caps, measuring the distance between the centre of plastids and morphological cell bottom. Considerable temporal differences have been determined between the kinetics of the longitudinal motion of amyloplasts in root and hypocotyl statocytes of 1-g seedlings upon exposition to fast clinorotation and inversion. In statocytes of both organs of seedlings grown under fast clinorotation, the gravity provoked displacement of statoliths in the direction of its action; however, the displacement was much faster in hypocotyl than in root statocytes. It has been assumed that the gravity-determined longitudinal transport of amyloplasts, both in hypocotyl endodermic cells and root statocytes of cress seedlings, is modulated by the cytoskeleton.     

Tubulin Cytoskeleton in Arabidopsis thaliana Root Cells Under Clinorotation

The effect of horizontal clinorotation (2 rpm) on the spatial organization of cortical microtubules (CMTs) in the living Arabidopsis root cells has been investigated. MTs were visualized by using a stably transformed line of transgenic Arabidopsis thaliana expressing GFP-MAP4 fusion protein. To monitor total changes in MT dynamics, the sensitivity to MT-inhibitor 5 μM/l oryzalin was used as an indicator. It was shown that CMTs in the distal elongation zone (DEZ) of clinorotated 5 day old seedlings exhibited a higher degree of disorder and also they were more sensitive to oryzalin. We suggest that rearrangement of CMTs in DEZ cells under clinorotation is caused by fast rate of MT dynamic turnover.     

Effect of Hypergravity on the Level of Heat Shock Proteins 70 and 90 in Pea Seedlings

Exposure to hypergravity induces significant changes in gene expression of plants which are indicative of stress conditions. A substantial part of the general stress response is up-regulation of heat shock proteins (Hsp) which function as molecular chaperones. The objective of this research was to test the possible changes in the Hsp70 and Hsp90 level in response to short-term hypergravity exposure. In this study 5-day-old etiolated pea seedlings were exposed to centrifuge-induced hypergravity (3–14 g) for 15 min and 1 h and a part of the seedlings was sampled at 1.5 and 24 h after the exposures. Western blot analysis showed time-dependent changes in Hsp70 and Hsp90 levels: an increase under hypergravity and a tendency towards recovery of the normal content during re-adaptation. The quantity and time of their expression was correlated with the g-force level. These data suggest that short-term hypergravity acts as a stress which could increase the risk of protein denaturation and aggregation. Molecular chaperons induced during the stress may have an essential role in counteracting this risk.     

Physiological Responses of Synechocystis sp. PCC 6803 under Clinorotation

Photosystem efficiency and the characteristic on oxidative stress were examined to elucidate the metabolic responses of Synechocystis sp. PCC 6803 to short-term clinorotation. Results compiled when using clinostat to simulate microgravity for 60?h, showed that clinorotation clearly prohibited the photochemical quantum yield, but promoted the synthesis of chlorophyll and total protein. This may be a compensatory mechanism for the algal cell to maintain its normal metabolism. An increased malondialdehyde (MDA) content of algal cell upon clinorotation, together with an enhanced catalase (CAT) activity was observed during the whole period of clinorotation. One conclusion is that short-term clinorotation acts as a kind of stress, and that these physiological responses may be a special way for an algal cell to adapt itself to a different environment other than earth gravity.     

Actin Microfilament Organization in the Transition Zone of Arabidopsis-ABD2-GFP Roots Under Clinorotation

Seedlings of Arabidopsis thaliana-ABD2-GFP were grown under slow clinorotation (2?rpm) and treated with actin and tubulin disrupting drugs in order to characterize the role of actin microfilaments in cell growth and gravisensing. Changes in microfilament organization and cell parameters have shown that the transition root zone (TZ) is rather sensitive to microfilament disruption in control plants. It is assumed that under clinorotation, organization of actin cytoskeleton in the TZ is coordinated in a different way than in the control. Organization of microfilaments depends upon organization of microtubules and clinorotation does not influence this interrelation significantly.     

Compressive pressure dependent anisotropic effective thermal conductivity of granular beds

In situ planetary thermal conductivity measurements are typically made using a long needle-like probe, which measures effective thermal conductivity in the probe??s radial (horizontal) direction. The desired effective vertical thermal conductivity for heat flow calculations is assumed to be the same as the measured effective horizontal thermal conductivity. However, it is known that effective thermal conductivity increases with increasing compressive pressure on granular beds and the horizontal stress in a granular bed under gravity is related to the vertical stress through Jaky??s at rest earth pressure coefficient. The objectives of this study were to examine the validity of the isotropic property assumption and to develop a fundamental understanding of the effective thermal conductivity of a dry, noncohesive granular bed under uniaxial compression. A model was developed to predict the increase in effective vertical and horizontal thermal conductivity with increasing compressive vertical applied pressure. An experiment was developed to simultaneously measure the effective vertical and horizontal thermal conductivities of particle beds with needle probes. Measurements were made as compressive vertical pressure was increased to show the relationship between increasing pressure and effective vertical and horizontal thermal conductivity. The results of this experiment showed quantitatively the conductivity anisotropy for two different materials and validated the developed model. This model can be used to predict the anisotropic effective thermal conductivity of granular materials under uniaxial compressive pressures, and evaluate the uncertainties in lunar heat flow measurements.     

Changes in Gene Expression of E. coli under Conditions of Modeled Reduced Gravity

Relatively few studies have examined bacterial responses to the reduced gravity conditions that are experienced by bacteria grown in space. In this study, whole genome expression of Escherichia coli K12 under clinorotation (which models some of the conditions found under reduced gravity) was analyzed. We hypothesized that phenotypic differences at cellular and population levels under clinorotation (hereafter referred to as modeled reduced gravity) are directly coupled to changes in gene expression. Further, we hypothesized that these responses may be due to indirect effects of these environmental conditions on nutrient accessibility for bacteria. Overall, 430 genes were identified as significantly different between modeled reduced gravity conditions and controls. Up-regulated genes included those involved in the starvation response (csiD, cspD, ygaF, gabDTP, ygiG, fliY, cysK) and redirecting metabolism under starvation (ddpX, acs, actP, gdhA); responses to multiple stresses, such as acid stress (asr, yhiW), osmotic stress (yehZYW), oxidative stress (katE, btuDE); biofilm formation (lldR, lamB, yneA, fadB, ydeY); curli biosynthesis (csgDEF), and lipid biosynthesis (yfbEFG). Our results support the previously proposed hypothesis that under conditions of modeled reduced gravity, zones of nutrient depletion develop around bacteria eliciting responses similar to entrance into stationary phase which is generally characterized by expression of starvation inducible genes and genes associated with multiple stress responses.     

Rocket seedling production on the international space station: Growth and nutritional properties

Producing sprouts directly during space missions may represent an interesting opportunity to offer high-quality fresh ready to eat food to the astronauts. The goal of this work was to compare, in terms of growth and nutritional quality, rocket (Eruca sativa Mill.) seedlings grown in the International Space Station during the ENEIDE mission with those grown in a ground-based experiment (in presence and absence of clinorotation). The rocket seedlings obtained from the space-experiment were thinner and more elongated than those obtained in the ground-based experiment. Cotyledons were often closed in the seedlings grown in the space experiment. Quantitative (germination, fresh and dry weight) and qualitative (glucose, fructose, sucrose and starch) traits of rocket seedling were negatively affected by micrograv-ity, especially those recorded on seedlings grown under real microgravity conditions The total chlorophyll, and carotenoids of seedlings obtained in the space experiment were strongly reduced in comparison to those obtained in the ground-based experiment (presence and absence of clinorotation). The results showed that it is possible to produce rocket seedlings in the ISS; however, further studies are needed to define the optimal environmental conditions for producing rocket seedlings with high nutritional value     

温度刺激下栉孔扇贝不同组织热休克蛋白HSP70的表达研究

采用免疫印迹(Western blot)和流式细胞术(flow cytometry,FCM)方法,研究了热激栉孔扇贝不同组织内热休克蛋白70(heat shock protein70,HSP70)的表达变化。免疫印迹结果显示,HSP70的诱导表达有组织特异性,即热休克可以使扇贝鳃组织中HSP70的表达明显升高,但在肌柱、外套膜和性腺组织中,未检测到HSP70的表达。流式细胞术结果表明：在不同的季节,相同的热刺激对扇贝血淋巴中HSP70的诱导表达存在差异。高温刺激后,栉孔扇贝血淋巴细胞中HSP70的表达量逐渐升高,在诱导7h时达到最高。     

Effects of Clinorotation on Growth and Chlorophyll Content of Rice Seeds

Microgravity, as a different environment, has been shown to affect plant growth and development (Sievers et al. 1996; Sack 1997). In the present study, effects of slow clinorotation (2 rpm) on growth and chlorophyll content in rice (variety: PRH-10) seedlings were investigated. Rice seeds were clinorotated continuously for 3, 5 and 7 days under ambient conditions. Root and shoot lengths and weights of rice seedlings were measured on the third, fifth and seventh day. Chlorophyll was extracted using N, N-Dimethylformamide (DMF). Absorption and fluorescence spectra of chlorophyll were recorded. Chlorophyll a, chlorophyll b and total chlorophyll contents were calculated from absorption spectra using Arnon’s method. Results showed an increase in root and shoot lengths in clinorotated samples. Similar results were obtained for root and shoot weights. Absorption spectra of chlorophyll showed no shift in the absorption peaks. Chlorophyll content was increased in clinorotated samples as compared to the controls. Interestingly, the difference between chlorophyll content in control and clinorotated samples decreased as the number of days of clinorotation increased. Chlorophyll a/b ratio was lowered in clinorotated samples as compared to the controls. These results suggest that slow clinorotation (2 rpm) affects plant growth and chlorophyll content in rice seedlings.     

输电塔线体系在摇摆地震动作用下的动力稳定性分析

为探究摇摆分量对输电塔线体系动力稳定性能的影响,以一实际输电塔为原型进行了动力时程分析,其中摇摆地震动采用改进的谱比法由地震平动中获取;根据增量动力分析(incremental dynamic analysis, IDA)方法结合B-R准则,分别对塔线体系在水平地震作用、水平-摇摆耦合地震作用和水平-竖向-摇摆耦合地震作用下的动力稳定性能进行分析。研究结果表明：摇摆分量及其引起的附加P-Δ效应会使输电塔线体系产生非对称振动,结构发生偏离平衡位置的单向偏移,从而导致塔线体系较仅考虑水平地震作用,更易动力失稳;重力和竖向地震响应下共同的二阶效应,放大了摇摆分量对结构动力稳定性能的影响,加剧输电塔线体系动力失稳破坏;发生动力失稳破坏时,薄弱区域主要集中于塔身中下部,杆件失效使塔身局部变形过大,导致塔线体系发生整体动力失稳。     

Cytosolic Calcium Concentration Changes in Neuronal Cells Under Clinorotation and in Parabolic Flight Missions

All life on earth has been established under conditions of stable gravity of 1g. Nevertheless, in numerous experiments the direct gravity dependence of biological processes has been shown on all levels of organization, from single molecules to humans. To study the effects especially of microgravity on biological systems, a variety of platforms are available, from drop towers to the ISS. Due to the costs of these platforms and their limited availability, as an alternative, numerous simulators have been developed for so called “simulated” microgravity. A classical systems is a clinostat, basically rotating a sample around one axis, and by integration of the gravity vector for 360° arguing that thus the effects of gravity are depleted. Indeed, a variety of studies has shown that taking out the direction of gravity from a biological system often results in consequences similar to the exposure of the system to real microgravity. Nevertheless, the opposite has been shown, too, and as a consequence the relevance of clinostats in microgravity research is still under discussion. To get some more insight into this problem we have constructed a small fluorescence clinostat and have studied the effects of clinorotation on the cytosolic calcium concentration of neuroglioma cells. The results have been compared to experiments with identical cells in real microgravity, utilizing parabolic flight missions. Our results show that in case of a cell suspension used in a small florescence clinostat within a tube diameter of 2mm, the effects of clinorotation are comparable to those under real microgravity, both showing a significant increase in intracellular calcium concentration.     

主动围压下岩石爆炸动态响应数值模拟

针对深部岩石在爆破作用下的失效机理和破坏过程,基于ANSYS/LSDYNA软件,在水平方向和竖直方向进行围压约束来模拟岩石受到的地应力作用。选取单向围压、等值双向围压和非等值双向围压对中心孔爆破下岩石动态响应进行研究。将数值模拟结果与模型实验结果对比,分析爆炸应力波的传播过程,并讨论了不同围压约束对爆炸后裂纹扩展的影响。结果表明,初始围压作用对爆炸应力波的传播影响较小;初始围压会抑制爆破裂纹扩张;竖直方向围压主要抑制水平方向裂纹扩张,水平方向围压主要抑制竖直方向裂纹扩张。     

主动围压下岩石爆炸动态响应数值模拟

针对深部岩石在爆破作用下的失效机理和破坏过程,基于ANSYS/LSDYNA软件,在水平方向和竖直方向进行围压约束来模拟岩石受到的地应力作用。选取单向围压、等值双向围压和非等值双向围压对中心孔爆破下岩石动态响应进行研究。将数值模拟结果与模型实验结果对比,分析爆炸应力波的传播过程,并讨论了不同围压约束对爆炸后裂纹扩展的影响。结果表明,初始围压作用对爆炸应力波的传播影响较小;初始围压会抑制爆破裂纹扩张;竖直方向围压主要抑制水平方向裂纹扩张,水平方向围压主要抑制竖直方向裂纹扩张。     

Investigation on Effect of Gravity Level on Bubble Distribution and Liquid Turbulence Modification for Horizontal Channel Bubbly Flow

Bubbly flows in the horizontal channel or pipe are often seen in industrial engineering fields, so it is very necessary to fully understand hydrodynamics of horizontal bubbly flows so as to improve industrial efficiency and to design an efficient bubbly system. In this paper, in order to fully understand mechanisms of phase distribution and liquid–phase turbulence modulation in the horizontal channel bubbly flow, the influence of gravity level on both of them were investigated in detail with the developed Euler–Lagrange two–way coupling method. For the present investigation, the buoyance on bubbles in both sides of the channel always points to the corresponding wall in order to study the liquid–phase turbulence modulation by bubbles under the symmetric physical condition. The present investigation shows that the gravity level has the important influence on the wall–normal distribution of bubbles and the liquid–phase turbulence modulation; the higher the gravity level is, the more bubbles can overcome the wall–normal resistance to accumulate near the wall, and the more obvious the liquid–phase turbulence modulation is. It is also discovered that interphase forces on the bubbles are various along the wall–normal direction, which leads to the fact that the bubble located in different wall–normal places has a different wall–normal velocity.     

多向地震耦合作用下高耸结构土-结构动力相互研究

研究了水平地震与竖向地震耦合作用下,考虑土-结构相互作用以及二阶效应对结构反应的影响。采用集总参数法,推导了结构在竖向地震和水平地震作用下的运动方程,对某高层建筑的地震反应进行了数值分析。结果表明:考虑土-结构相互作用使结构周期延长,变形增大;竖向地震作用使结构的地震反应有不同程度的增加;考虑二阶效应后结构的位移反应以及层间剪力增加。故在设计建筑在软土地基上的高层及高耸结构时,应该考虑结构的重力二阶效应,考虑其对结构的整体稳定性的不利影响。     

Clinorotation Affects the Ultrastructure of Pea Root Mitochondria

Effects of clinorotation on the mitochondrial ultrastructure in cells of meristem, distal and central elongation zones of 3- and 5-day-old etiolated roots of pea seedling roots were studied. It was shown that mitochondria in cells of examined root growth zones revealed a different sensitivity to clinorotation. The ultrastructure of mitochondria in the meristem and central elongation zone cells did not substantially change in comparison with stationary control. At the same time, changes in the mitochondrial ultrastructure in cells of the distal elongation zone under clinorotation were observed, namely: decrease in the size of mitochondria, as well as increase in both matrix electron density and crista volume. Such changes in the mitochondrial ultrastructure under clinorotation are supposed to display the rearrangements of energy metabolism in cells of the distal elongation zone in these conditions.