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.     

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.     

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.     

The stress protein level under clinorotation in context of the seedling developmental program and the stress response

Heat-shock proteins (HSP70 and HSP90) are present in plant cells under the normal growth conditions. At the same time, a variety of environmental disruptions results in their rapid synthesis as a substantial part of adaptation. HSP amounts can be indicative of a cellular stress level. Altered gravity (clinorotation) is unnatural for plants, so it may be a kind of stress. The aim of this study was to analyze the influence of horizontal clinorotation on the HSP70 and HSP90 level during seedling development. Pea (Pisum sativum L.) seedlings grown for 3 days from seed imbibitions in stationary control and under slow clinorotation (2 rpm) are used for this investigation. Western blot analysis indicated that HSP70 and HSP90 were abundant in the embryos of dry seeds and their amount decreased significantly during seed germination. But under horizontal clinorotation, their level in seedlings remained higher compared to the control. Furthermore, a comparison of the influence of horizontal and vertical clinorotation on the HSP level was carried out. On the ELISA data, HSP70 and HSP90 amounts in the 3-day old seedlings were higher after horizontal clinorotation than after vertical. The obtained data show an increased HSP70 and HSP90 level in pea seedlings under clinorotation. Both, rotation and change in the cell position relatively to a gravity vector affect the HSP level.     

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.     

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     

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.     

Analysis of Statoliths Displacement in <Emphasis Type="Italic">Chara</Emphasis> Rhizoids for Validating the Microgravity-Simulation Quality of Clinorotation Modes

In single-celled rhizoids of the green algae Chara, positively gravitropic growth is governed by statoliths kept in a dynamically stable position 10–25 μ m above the cell tip by a complex interaction of gravity and actomyosin forces. Any deviation of the tube-like cells from the tip-downward orientation causes statoliths to sediment onto the gravisensitive subapical cell flank which initiates a gravitropic curvature response. Microgravity experiments have shown that abolishing the net tip-directed gravity force results in an actomyosin-mediated axial displacement of statoliths away from the cell tip. The present study was performed to critically assess the quality of microgravity simulation provided by different operational modes of a Random Positioning Machine (RPM) running with one axis (2D mode) or two axes (3D mode) and different rotational speeds (2D), speed ranges and directions (3D). The effects of 2D and 3D rotation were compared with data from experiments in real microgravity conditions (MAXUS sounding rocket missions). Rotational speeds in the range of 60–85 rpm in 2D and 3D modes resulted in a similar kinetics of statolith displacement as compared to real microgravity data, while slower clinorotation (2–11 rpm) caused a reduced axial displacement and a more dispersed arrangement of statoliths closer to the cell tip. Increasing the complexity of rotation by adding a second rotation axis in case of 3D clinorotation did not increase the quality of microgravity simulation, however, increased side effects such as the level of vibrations resulting in a more dispersed arrangement of statoliths. In conclusion, fast 2D clinorotation provides the most appropriate microgravity simulation for investigating the graviperception mechanism in Chara rhizoids, whereas slower clinorotation speeds and rotating samples around two axes do not improve the quality of microgravity simulation.     

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.     

Effect of carbon nanomaterials on the germination and growth of rice plants

For the successful diverse applications of different nanomaterials in life sciences, it is necessary to understand the ultimate fate, distribution and potential environmental impacts of manufactured nanomaterials. Phytotoxicity studies using higher plants is an important criterion for understanding the toxicity of engineered nanomaterials. We studied the effects of engineered carbon nanomaterials of various dimensionalities (carbon nanotubes, C60, graphene) on the germination of rice seeds. A pronounced increase in the rate of germination was observed for rice seeds in the presence of some of these carbon nanostructures, in particular the nanotubes. Increased water content was observed in the carbon nanomaterial treated seeds during germination compared to controls. The germinated seeds were then grown in a basal growth medium supplemented with carbon nanomaterials for studying their impact on further seedling growth. Treated seedlings appeared to be healthier with well-developed root and shoot systems compared to control seedlings. Our results indicate the possible use for carbon nanomaterials as enhancers in the growth of rice seedlings.     

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.     

Modelling colour and chlorophyll losses of frozen green beans (Phaseolus vulgaris, L.)

Colour changes and chlorophyll degradation of frozen green beans (Phaseolus vulgaris, L., variety bencanta) were studied during 250 days of storage at −7, −15 and −30 °C. Chlorophyll a and b losses and colour Hunter a and b co-ordinates and total colour difference (TCD_H) changes were successfully described by first order and reversible first order models, respectively. The temperature effect was described by the Arrhenius law. Disagreement between the colour co-ordinates and chlorophyll content was obtained. Therefore, chlorophyll content is not a good colour index of frozen green beans. The results emphasise that colour is a more important parameter to assess frozen green beans visual quality.     

Deep-level studies in GaN layers grown by epitaxial lateral overgrowth

The electrical properties, deep-level spectra, microcathodoluminescence (MCL) spectra and diffusion lengths of minority charge carriers were measured in GaN films grown by the epitaxial lateral overgrowth (ELOG) technique. The results are compared to the properties of GaN layers grown in a standard fashion without masking of the initial template. MCL and electron beam induced current (EBIC) imaging of the laterally overgrown regions revealed the presence of dark spots with density of 1-5 × 10⁶ cm^− 2 that are associated with individual dislocations. The concentration of deep electron and hole traps was found to be much higher in the standard material than in the ELOG material. Diffusion lengths of minority carriers determined from EBIC signal profiling gave values of 0.8-1 μm along the bright regions and 0.4-0.5 μm in the dark regions of the ELOG samples. Similar measurements on metal organic chemical vapor deposition templates gave a diffusion length of 0.4-0.5 μm, close to the diffusion length in the dark stripes of the ELOG samples.     

Grinding effects on the change of particle properties in cupric sulfide,CuS

Cupric sulfide, CuS, was ground by a planetary ball mill under various experimental conditions, including variations in rotation speed and grinding time. The changes in crystal structure, lattice parameter, particle size, pore structure, and specific surface area in the ground powders were determined. With regard to the CuS ground at 700 rpm, the changes of the crystal structure were more remarkable than those of the CuS ground at 300 rpm. The crystallite size decreased (from 18.5 to 8.4 nm) and the lattice strain increased (from 1.60% to 3.09%) in the CuS ground at 700 rpm. Moreover, the lattice parameter a increased, while the lattice parameter c decreased. Grinding at 300 rpm is favorable for inducing changes of the physical properties, such as the growth of pore volume and specific surface area.     

Fish Inner Ear Otolith Growth Under Real Microgravity (Spaceflight) and Clinorotation

Using late larval stages of cichlid fish (Oreochromis mossambicus) we have shown earlier that the biomineralization of otoliths is adjusted towards gravity by means of a neurally guided feedback loop. Centrifuge experiments, e.g., revealed that increased gravity slows down otolith growth. Microgravity thus should yield an opposite effect, i.e., larger than normal otoliths. Consequently, late larval cichlids (stage 14, vestibular system operational) were subjected to real microgravity during the 12 days FOTON-M3 spaceflight mission (OMEGAHAB-hardware). Controls were kept at 1g on ground within an identical hardware. Animals of another batch were subsequently clinorotated within a submersed fast-rotating clinostat with one axis of rotation (2d-clinostat), a device regarded to simulate microgravity. Temperature and light conditions were provided in analogy to the spaceflight experiment. Controls were maintained at 1g within the same aquarium. After all experiments, animals had reached late stage 21 (fish can swim freely). Maintenance under real microgravity during spaceflight resulted in significantly larger than normal otoliths (both lapilli and sagittae, involved in sensing gravity and the hearing process, respectively). This result is fully in line with an earlier spaceflight study in the course of which otoliths from late-staged swordtails Xiphophorus helleri were analyzed. Clinorotation resulted in larger than 1g sagittae. However, no effect on lapilli was obtained. Possibly, an effect was present but too light to be measurable. Overall, spaceflight obviously induces an adaptation of otolith growth, whereas clinorotation does not fully mimic conditions of microgravity regarding late larval cichlids.     

Strategies of biosynthesis of poly(3-hydroxybutyrate) supplemented with biodiesel obtained from rice bran oil

The persistence of conventional synthetic plastic in the environment is increasingly considered as a source of ecological problem and has been motivated the research of biodegradable and biocompatible polymers. Polyhydroxyalkanoates (PHAs) are thermoplastic polyesters synthesized by several bacteria and Cupriavidus necator is the known one. Due to the high production cost of PHAs, the use of a cheaper carbon source or nutritional supplements are required in order to reduce the production cost. The aim of this research was to investigate the influence of biodiesel of rice bran oil as a low cost nutritional supplement in cultures of C. necator in mineral medium on the cell growth and production of poly(3-hydroxybutyrate) - P(3HB). For this, C. necator was cultivated in shake flasks of 500 mL with 150 mL of mineral medium at 30 °C for 24 h in 150 rpm. Samples were removed each 4 h for evaluation. The results demonstrate that the addition of biodiesel of rice bran oil as nutritional supplement in the culture of C. necator was viable, since its addition allows a cell growth with substrate consume reduction. Besides, the P(3HB) yield was 61% and 150% superior when biodiesel was used compared to the control medium using 30 g L^? 1 and 15 g L^? 1, respectively.     

Effect of welding parameters on microstructure and mechanical properties of friction stir welded joints of a super high strength Al–Zn–Mg–Cu aluminum alloy

Samples made of a super high strength aluminum alloy with high Zn content were friction stir welded with rotation rates of 350–950 rpm and welding speeds of 50–150 mm/min. The effect of welding parameters on the microstructure and mechanical properties was investigated. It was observed that the grain size of the nugget zones decreased with the increasing welding speed or the decreasing tool rotation rate. Most of the strengthening precipitates in the nugget zone were dissolved back and the intragranular and grain boundary precipitates in the heat affected zone coarsened significantly. The greatest ultimate tensile strength of 484 MPa and largest elongation of 9.4 were obtained at 350 rpm−100 mm/min and 350 rpm−50 mm/min, respectively. The ultimate tensile strength and elongation deteriorated drastically when rotation rate increased from 350 to 950 rpm at a constant welding speed of 100 mm/min.     

Spectroscopic study of Dy and Dy/Yb ions co-doped in barium fluoroborate glass

Dy³⁺ and Dy³⁺/Yb³⁺ co-doped barium fluoroborate glasses have been synthesized and their luminescence properties have been monitored under UV (355 nm) and NIR (976 nm) excitations, respectively. Absorption spectra of the two samples were recorded and Judd–Ofelt intensity parameters and other radiative parameters were calculated and compared. Dy³⁺ doped sample yields strong yellowish white emission under 355 nm excitation. No emission is observed with 976 nm excitation. The Dy³⁺/Yb³⁺ ions co-doped sample on the other hand gives weak upconversion emission in blue, yellow and red regions on 976 nm excitation via energy transfer process from Yb³⁺ to Dy³⁺ ions from other level. An involvement of three photons has been noted for these emissions. Decay curve for different transitions of these samples have also been recorded and the lifetime of the corresponding levels involved are calculated.     

Antireflection and band gap extension effects of ZnO nanocrystalline films grown on ITO-coated glasses by low temperature process

Zinc oxide (ZnO) nanocrystalline films coated on indium tin oxides (ITO, 90:10 wt%) glasses were prepared by low temperature process. The thin films were composed of uniform nanoparticles with average diameter around 8.4 nm. All samples exhibited excellent optical antireflective phenomena, and the maximum transmission reached 92.8% for the sample spin coated at 1500 rpm at 453 nm, improved by 21.5%. The antireflective results were explained by the coherence theory. And the antireflective effects were induced by the ITO and ZnO films. The calculated thicknesses of the ZnO films agreed well with the experimental results. The theoretical calculated band gap from the average diameter of ZnO nanoparticles was also well consistent with the experimental ones obtained from the optical transmission spectra. This result was promising for applications in organic solar cells.