Leaf senescence under various gravity conditions: relevance to the dynamics of plant hormones.

Effects of simulated microgravity and hypergravity on the senescence of oat leaf segments excised from the primary leaves of 8-d-old green seedlings were studied using a 3-dimensional (D) clinostat as a simulator of weightlessness and a centrifuge, respectively. During the incubation with water under 1-g conditions at 25 degrees C in the dark, the loss of chlorophyll of the segments was found dramatically immediately after leaf excision, and leaf color completely turned to yellow after 3-d to 4-d incubation. In this case kinetin (10 micromolar) was effective in retarding senescence. The application of simulated microgravity conditions on a 3-D clinostat enhanced chlorophyll loss in the presence or absence of kinetin. The loss of chlorophyll was also enhanced by hypergravity conditions (ca. 8 to 16 g), but the effect was smaller than that of simulated microgravity conditions on the clinostat. Jasmonates (JAs) and abscisic acid (ABA) promoted senescence under simulated microgravity conditions on the clinostat as well as under 1-g conditions. After 2-d incubation with water or 5-d incubation with kinetin, the endogenous levels of JAs and ABA of the segments kept under simulated microgravity conditions on the clinostat remained higher than those kept under 1-g conditions. These findings suggest that physiological processes of leaf senescence and the dynamics of endogenous plant hormone levels are substantially affected by gravity.     

The effect of gravity on plant germination.

An axis clinostat was constructed to create micro and negative gravity also a rotated flat disk was constructed with different rotation rates to give increased gravity, by centrifugal force up to 48 g. Rice seeds were grown on agar in tubes at the constant air temperature of 20 degrees C under an average light condition of 110 micromol/m2/sec(PPF). Humidity was not controlled but was maintained above 90%. Since the tube containers were not large enough for long cultivation, shoot and root growth were observed every 12 hours until the sixth day from seeding. The lengths of shoots and roots for each individual plant were measured on the last day. The stem lengths were increased by microgravity but the root lengths were not. Under the negative gravity, negative orthogeotropism and under microgravity, diageotropism was observed. No significant effect of increased gravity was observed on shoot and root growth.     

Effect of the space environment on the induction of DNA-repair related proteins and recovery from radiation damage.

Recovery of bacterial cells from radiation damage and the effects of microgravity were examined in an STS-79 Shuttle/Mir Mission-4 experiment using the extremely radioresistant bacterium Deinococcus radiodurans. The cells were irradiated with gamma rays before the space flight and incubated on board the Space-Shuttle. The survival of the wild type cells incubated in space increased compared with the ground controls, suggesting that the recovery of this bacterium from radiation damage was enhanced under microgravity. No difference was observed for the survival of radiosensitive mutant rec30 cells whether incubated in space or on the ground. The amount of DNA-repair related RecA protein induced under microgravity was similar to those of ground controls, however, induction of PprA protein, the product of a newly found gene related to the DNA repair mechanism of D. radiodurans, was enhanced under microgravity compared with ground controls.     

The effects of microgravity on induced mutation in Escherichia coli and Saccharomyces cerevisiae.

We examined whether microgravity influences the induced-mutation frequencies through in vivo experiments during space flight aboard the space shuttle Discovery (STS-91). We prepared dried samples of repair-deficient strains and parental strains of Escherichia (E.) coli and Saccharomyces (S.) cerevisiae given DNA damage treatment. After culture in space, we measured the induced-mutation frequencies and SOS-responses under microgravity. The experimental findings indicate that almost the same induced-mutation frequencies and SOS-responses of space samples were observed in both strains compared with the ground control samples. It is suggested that microgravity might not influence induced-mutation frequencies and SOS-responses at the stages of DNA replication and/or DNA repair. In addition, we developed a new experimental apparatus for space experiments to culture and freeze stocks of E. coli and S. cerevisiae cells.     

Effects of several environmental factors on sweetpotato growth.

Effects of relative humidity, light intensity and photoperiod on growth of 'Ga Jet' and 'TI-155' sweetpotato cultivars, using the nutrient film technique (NFT), have been reported. In this study, the effect of ambient temperature regimes (constant 28 degrees C and diurnal 28:22 degrees C day:night) and different CO2 levels (ambient, 400, 1000 and 10000 microliters/L--400, 1000 and 10000 ppm) on growth of one or both of these cultivars in NFT are reported. For a 24-h photoperiod, no storage roots were produced for either cultivar in NFT when sweetpotato plants were grown at a constant temperature of 28 degrees C. For the same photoperiod, when a 28:22 degrees C diurnal temperature variation was used, there were still no storage roots for 'TI-155' but the cv. 'Ga Jet' produced 537 g/plant of storage roots. For both a 12-h and 24-h photoperiod, 'Ga Jet' storage root fresh and dry weight tended to be higher with a 28:22 degrees C diurnal temperature variation than with a constant 28 degrees C temperature regime. Preliminary results with both 'Ga Jet' and 'TI 155' cultivars indicate a distinctive diurnal stomatal response for sweetpotato grown in NFT under an ambient CO2 level. The stomatal conductance values observed for 'Ga Jet' at elevated CO2 levels indicated that the difference between the light- and dark-period conductance rates persisted at 400, 1000, and 10000 microliters/L.     

Evaluation of using two-phase frictional pressure drop correlations for normal gravity to microgravity and reduced gravity

The calculation of two-phase frictional pressure drop (TPFPD) is required by two-phase systems operating under microgravity and reduced gravity. There are a large number of correlations for the TPFPD in tubes under normal gravity. However, it is hard to find out a TPFPD correlation obtained from microgravity and/or reduced gravity conditions, and thus people have to use TPFPD correlations for normal gravity to calculate TPFPD under microgravity and reduced gravity. It is necessary to evaluate the feasibility of such practice. This paper offers a comprehensive review of the TPFPD correlations for normal gravity and an up-to-data survey of the TPFPD experimental study under microgravity and reduced gravity. There are 23 TPFPD correlations for normal gravity reviewed and 135 experimental data under microgravity obtained from the literature. These experimental data are used to evaluate the reviewed TPFPD correlations. It is found that the smallest mean absolute relative deviation (MARD) of the correlations is greater than 34%. Using TPFPD correlations for normal gravity to reduced gravity and microgravity may be acceptable for the first approximation, but correlations intended for microgravity and reduced gravity are needed and more experiments are desired to obtain more data with high accuracy.     

Gravitational force regulates elongation growth of Arabidopsis hypocotyls by modifying xyloglucan metabolism.

Growth of dark-grown Arabidopsis hypocotyls was suppressed under hypergravity conditions (300 g), or was stimulated under microgravity conditions in space (Space Shuttle STS-95). The mechanical extensibility of cell walls decreased and increased under hypergravity and microgravity conditions, respectively. The amounts of cell wall polysaccharides (pectin, hemicellulose-I, hemicellulose-II and cellulose) per unit length of hypocotyls increased under hypergravity conditions, and decreased under microgravity conditions. The amount and the molecular mass of xyloglucans also increased under the hypergravity conditions, while those decreased under microgravity conditions. The activity of xyloglucan-degrading enzymes extracted from hypocotyl cell walls decreased and increased under hypergravity and microgravity conditions, respectively. These results indicate that the amount and the molecular mass of xyloglucans are affected by the magnitude of gravity and that such changes are caused by changes in xyloglucan-degrading activity. Modifications of xyloglucan metabolism as well as the thickness of cell walls by gravity stimulus may be the primary event determining the cell wall extensibility, thereby regulating the growth rate of Arabidopsis hypocotyls.     

Preliminary results of space experiment: Implications for the effects of space radiation and microgravity on survival and mutation induction in human cells

In view of the concern for the health of astronauts that may one day journey to Mars or the Moon, we investigated the effect that space radiation and microgravity might have on DNA damage and repair. We sent frozen human lymphoblastoid TK6 cells to the International Space Station where they were maintained under frozen conditions during a 134-day mission (14 November 2008 to 28 March 2009) except for an incubation period of 8 days under 1G or μG conditions in a CO₂ incubator. The incubation period started after 100 days during which the cells had been exposed to 54 mSv of space radiation. The incubated cells were then refrozen, returned to Earth, and compared to ground control samples for the determination of the influence of microgravity on cell survival and mutation induction. The results for both varied from experiment to experiment, yielding a large SD, but the μG sample results differed significantly from the 1G sample results for each of 2 experiments, with the mean ratio of μG to 1G being 0.55 for the concentration of viable cells and 0.59 for the fraction of thymidine kinase deficient (TK⁻) mutants. Among the mutants, non-loss of zygosity events (point mutations) were less frequent (31%) after μG incubation than after 1G incubation, which might be explained by the influence of μG on cellular metabolic or physiological function. Additional experiments are needed to clarify the effect of μG interferes on DNA repair.     

Water movement on the convex surfaces of porous media under microgravity

A plant growth system for crop production under microgravity is part of a life supporting system designed for long-duration space missions. A plant growth in soil in space requires the understanding of water movement in soil void spaces under microgravity. Under 1G-force condition, on earth, water movement in porous media is driven by gradients of matric and gravitational potentials. Under microgravity condition, water movement in porous media is supposed to be driven only by a matric potential gradient, but it is still not well understood. We hypothesized that under microgravity water in void spaces of porous media hardly moved comparing in void spaces without obstacles because the concave surfaces of the porous media hindered water movement. The objective of this study was to investigate water movement on the convex surfaces of porous media under microgravity. We conducted parabolic flight experiments that provided 20–25?s of microgravity at the top of a parabolic flight. We observed water movement in void spaces in soil-like porous media made by glass beads and glass spheres (round-bottomed glass flasks) in the different conditions of water injection under microgravity. Without water injection, water did not move much in neither glass beads nor glass spheres. When water was injected during microgravity, water accumulated in contacts between the particles, and the water made thick fluid films on the particles surface. When the water injection was stopped under microgravity, water was held in the contacts between the particles. This study showed that water did not move upward in the void spaces with or without the water injection. In addition, our results suggested that the difficulty of water movement on the convex (i.e. particle surfaces) might result in slower water move in porous media under microgravity than at 1G-force.     

重力对单气泡生长特性的影响

采用VOF (Volume of Fluid)多相流模型, 通过用户自定义函数UDF (User Defined Function)实现相变过程中质量和能量的输运, 对微重力条件下尺寸为10mm × 10mm × 25mm的矩形通道的池沸腾现象进行数值模拟, 得到了微重力及常重力作用下单个气泡生长特性的差异. 模拟结果表明, 微重力条件下气泡周围的流线与温度场的分布有显著差异; 由于表面张力作用, 微重力下的气泡脱离特性与常重力下不同; 在微重力条件下, 气泡直径的变化比较复杂, 并与重力加速度的大小有关; Marangoni流对微重力下的流动影响很大, 使换热系数波动, 而且波动的幅度随重力加速度的减小而增大.      

Utilization of white potatoes in CELSS.

Potatoes (Solanum tuberosum) have a strong potential as a useful crop species in a functioning CELSS. The cultivar Denali has produced 37.5 g m-2 d-1 when grown for 132 days with the first 40 days under a 12-h photoperiod and a light:dark temperature cycle of 20 degrees C:16 degrees C, and then 92 days under continuous irradiance and a temperature of 16 degrees C. Irradiance was at 725 micromoles m-2 s-1 PPF and carbon dioxide at 1000 micromoles mol-1. The dried tubers had 82% carbohydrates, 9% protein and 0.6% fat. Other studies have shown that carbon dioxide supplementation (1000 micromoles mol-1) is of significant benefit under 12-h irradiance but less benefit under 24 h irradiance. Irradiance cycles of 60 minutes light and 30 minutes dark caused a reduction of more than 50% in tuber weight compared to cycles of 16 h light and 8 h dark. A diurnal temperature change of 22 degrees C for the 12-h light period to 14 degrees C during the 12-h dark period gave increased yields of 30% and 10% for two separate cultivars, compared with plants grown under a constant 18 degrees C temperature. Cultivar screening under continuous irradiance and elevated temperatures (28 degrees C) for 8 weeks of growth indicated that the cvs Haig, Denali, Atlantic, Desiree and Rutt had the best potential for tolerance to these conditions. Harvesting of tubers from plants at weekly intervals, beginning at 8 weeks after planting, did not increase yield over a single final harvest. Spacing of plants on 0.055 centers produced greater yield per m2 than spacing at 0.11 or 0.22 m2. Plants maintained 0.33 meters apart (0.111 m2 per plant) in beds produced the same yields when separated by dividers in the root matrix as when no separation was made.     

Kinetics of amyloplast movement in cress root statocytes under different gravitational loads.

In order to investigate the movement of a statolith complex along the longitudinal axis of root cap statocytes under different mass accelerations, a series of experiments with Lepidium sativum L. in an automatically operating centrifuge during the Bion-11 satellite flight and on a centrifuge-clinostat have been performed. During spaceflight, roots were grown for 24 h under root-tip-directed centrifugal 1-g acceleration, then exposed to microgravity for 6, 12 and 24 min and chemically fixed. During the first 6 min of microgravity, the statoliths moved towards the cell center with a mean velocity of 0.31 +/- 0.04 micrometers/min, which decreased to 0.12 +/- 0.01 micrometers/min within subsequent 12-24 min period. The mean relative position of the statolith complex in respect to the distal cell wall (% of total cell length) increased from 24.0 +/- 0.5% in 1 g-grown roots to 38.8 +/- 0.8% in roots exposed for 24 min to microgravity, but remained smaller than in roots grown continuously in microgravity (48.0 +/- 0.7%). The properties of the statolith movement away from the distal pole of the statocyte were studied in roots grown for 24 h vertically under 1 g and then placed for 6 min on a fast rotating clinostat (50 rpm) or 180 degrees inverted. After 2 min of both treatments, the mean relative position of the statoliths increased by about 10% versus its initial position. Later on, the proximal displacement of amyloplasts slowed down under simulated weightlessness, while it proceeded at a constant velocity under 1 g inversion. In roots grown on the clinostat and then exposed to 1 g in the longitudinal direction, amyloplast sedimentation away from the central region of statocyte was similar at the beginning of distal and proximal 6-min 1-g stimulation. However, at the end of this period statolith displacement was more pronounced in proximal direction as compared to distal. It is proposed that statolith position in the statocyte of a vertical root is controlled by the force of gravity, however, the intracellular forces, first of all those generated by the network of the cytoskeleton, are manifested when an usual orientation of the organ is changed or the statocytes are exposed to microgravity and clinorotation.     

Embryogenesis and organogenesis of Carausius morosus under spaceflight conditions

The influence of cosmic radiation and/or microgravity on insect development was studied during the 7 day German Spacelab Mission D1. Eggs of Carausius morosus of five stages differing in sensitivity to radiation and in capacity to regeneration were allowed to continue their development in the BIORACK 22°C incubator, either at microgravity conditions or on the 1 g reference centrifuge. Using the Biostack concept - eggs in monolayers were sandwiched between visual track detectors - and the 1 g reference centrifuge, we were able to separate radiation effects from microgravity effects and also from combined effects of these two factors in space. After retrieval, hatching rates, growth kinetics and anomaly frequencies were determined in the different test samples. The early stages of development turned out to be highly sensitive to single hits of cosmic ray particles as well as to the temporary exposure to microgravity during their development. In some cases, the combined action of radiation and microgravity even amplified the effects exerted by the single parameters of space. Hits by single HZE particles caused early effects, such as body anomalies, as well as late effects, such as retarded growth after hatching. Microgravity exposure lead to a reduced hatching rate. A synergistic action of HZE particle hits and microgravity was established in the unexpectedly high frequency of anomal larvae. However, it cannot be excluded, that cosmic background radiation or low LET HZE particles are also causally involved in damage observed in the microgravity samples.     

微重力下胶原蛋白纤维化及羟基磷灰石结晶的初步研究

在长期空间飞行过程中, 骨质丢失是一个严重问题. 羟基磷灰石(HAP)晶体是骨骼的主要成分, 骨骼中的胶原蛋白纤维在HAP生长结晶过程中起到关键作用. 研究了胶原蛋白纤维化过程在模拟微重力和常重力条件下的变化, 对以胶原 蛋白纤维作为模板生长出的HAP晶体形貌进行了观察. 结果表明, 不同浓度胶原蛋白溶液中形成的胶原蛋白纤维, 其内部孔隙数量和尺寸在模拟微重力条件下要明显大于常重力条件下, 胶原蛋白纤维内部孔隙的分布也不同于常重力条 件下的结果. 以模拟微重力条件下形成的胶原蛋白纤维为模板生长出的HAP 晶体主要为立方体状, 而以常重力条件下形成的胶原蛋白纤维为模板生长出的 HAP晶体形貌主要为板状. 该结果有助于未来进一步阐明空间骨质丢失的机理.      

The effect of gravity on surface temperature and net photosynthetic rate of plant leaves.

To clarify the effects of gravity on heat/gas exchange between plant leaves and the ambient air, the leaf temperatures and net photosynthetic rates of plant leaves were evaluated at 0.01, 1.0, 1.5 and 2.0 G of 20 seconds each during a parabolic airplane flight. Thermal images of leaves were captured using infrared thermography at an air temperature of 26 degrees C, a relative humidity of 15% and an irradiance of 260 W m-2. The net photosynthetic rates were determined by using a chamber method with an infrared gas analyzer at an air temperature of 20 degrees C, a relative humidity of 50% and a photosynthetic photon flux of 0.5 mmol m-2 s-1. The mean leaf temperature increased by 1 degree C and the net photosynthetic rate decreased by 13% with decreasing gravity levels from 1.0 to 0.01 G. The leaf temperature decreased by 0.5 degree C and the net photosynthetic rate increased by 7% with increasing gravity levels from 1.0 to 2.0 G. Heat/gas exchanges between leaves and the ambient air were more retarded at lower gravity levels. A restricted free air convection under microgravity conditions in space would limit plant growth by retarding heat and gas exchanges between leaves and the ambient air.     

Effects of microgravity on c-fos gene expression in osteoblast-like MC3T3-E1 cells.

The paper summarizes the data on proliferation and gravity-related gene expression of osteoblasts that were obtained from an experiment conducted under simulated and real microgravity conditions. Simulated microgravity conditions obtained in a clinostat depress proliferation of both osteoblast-like MC3T3-E1 and HeLa carcinoma cells. This depression of proliferation occurs in a collagen gel culture in which the flow of culture medium by rotation may be reduced. Interestingly, MC3T3-E1 cells which are probably one of target cells to microgravity are more sensitive than the HeLa cells. Simulated microgravity inhibited the epidermal growth factor (EGF)-induced c-fos gene expression in the MC3T3-El cells. To examine in detail the effect of real microgravity on the EGF signal transduction cascade in osteoblasts, MC3T3-E1 cells were cultured in the Cell Culture Experiment Module of the sounding rocket TR-1A6. The EGF-induced c-fos expression in cells was depressed under short-term microgravity conditions in the sounding rocket, while the phosphorylation of mitogen-activated protein kinase (MAPK) was not affected compared with the controls grown on the ground. These results suggest that an action site of microgravity in the signal transduction pathway may be downstream of MAPK.     

Effects of lighting and air movement on temperatures in reproductive organs of plants in a closed plant growth facility

Temperature increases in plant reproductive organs such as anthers and stigmas could cause fertility impediments and thus produce sterile seeds under artificial lighting conditions without adequately controlled environments in closed plant growth facilities. There is a possibility such a situation could occur in Bioregenerative Life Support Systems under microgravity conditions in space because there will be little natural convective or thermal mixing. This study was conducted to determine the temperature of the plant reproductive organs as affected by illumination and air movement under normal gravitational forces on the earth and to make an estimation of the temperature increase in reproductive organs in closed plant growth facilities under microgravity in space. Thermal images of reproductive organs of rice and strawberry were captured using infrared thermography at air temperatures of 10–11 °C. Compared to the air temperature, temperatures of petals, stigmas and anthers of strawberry increased by 24, 22 and 14 °C, respectively, after 5 min of lighting at an irradiance of 160 W m⁻² from incandescent lamps. Temperatures of reproductive organs and leaves of strawberry were significantly higher than those of rice. The temperatures of petals, stigmas, anthers and leaves of strawberry decreased by 13, 12, 13 and 14 °C, respectively, when the air velocity was increased from 0.1 to 1.0 ms⁻¹. These results show that air movement is necessary to reduce the temperatures of plant reproductive organs in plant growth facilities.     

Volatile metabolites of higher plant crops as a photosynthesizing life support system component under temperature stress at different light intensities.

The effect of elevated temperatures of 35 and 45 degrees C (at the intensities of photosynthetically active radiation 322, 690 and 1104 micromoles m-2 s-1) on the photosynthesis, respiration, and qualitative and quantitative composition of the volatiles emitted by wheat (Triticum aestuvi L., cultivar 232) crops was investigated in growth chambers. Identification and quantification of more than 20 volatile compounds (terpenoids--alpha-pinene, delta 3 carene, limonene, benzene, alpha- and trans-caryophyllene, alpha- and gamma-terpinene, their derivatives, aromatic hydrocarbons, etc.) were conducted by gas chromatograph/mass spectrometry. Under light intensity of 1104 micromoles m-2 s-1 heat resistance of photosynthesis and respiration increased at 35 degrees C and decreased at 45 degrees C. The action of elevated temperatures brought about variations in the rate and direction of the synthesis of volatile metabolites. The emission of volatile compounds was the greatest under a reduced irradiation of 322 micromoles m-2 s-1 and the smallest under 1104 micromoles m-2 s-1 at 35 degrees C. During the repair period, the contents and proportions of volatile compounds were different from their initial values, too. The degree of disruption and the following recovery of the functional state depended on the light intensity during the exposure to elevated temperatures. The investigation of the atmosphere of the growth chamber without plants has revealed the substances that were definitely technogenic in origin: tetramethylurea, dimethylsulfide, dibutylsulfide, dibutylphthalate, and a number of components of furan and silane nature.     

大鼠后肢去负荷体位调节装置设计与实验研究

通过对大鼠尾吊模型进行改进，研制出一种新型可调节体位的大鼠后肢去负荷悬吊装置，研究模拟微重力效应下体液分布变化对大鼠骨代谢的影响.将36只SD大鼠均分为对照组（CON）、头低位后肢去负荷组（HDT）、水平位后肢去负荷组（HH）和头高位后肢去负荷组（HUT）4组，实验21天后，利用DXA检测大鼠的骨密度（BMD）.模拟微重力效应下的三组大鼠后肢均发生严重骨丢失，其中HH和HUT组后肢BMD显著大于HDT组.实验结果表明，体液分布变化可能在模拟微重力效应导致的骨丢失中起到重要作用，新型大鼠后肢去负荷悬吊装置能够调节大鼠体位（体液）进行模拟微重力效应研究.      

Function of the cytoskeleton in gravisensing during spaceflight.

Since astronauts and cosmonauts have significant bone loss in microgravity we hypothesized that there would be physiological changes in cellular bone growth and cytoskeleton in the absence of gravity. Investigators from around the world have studied a multitude of bone cells in microgravity including Ros 17/2.8, Mc3T3-E1, MG-63, hFOB and primary chicken calvaria. Changes in cytoskeleton and extracellular matrix (ECM) have been noted in many of these studies. Investigators have noted changes in shape of cells exposed to as little as 20 seconds of microgravity in parabolic flight. Our laboratory reported that quiescent osteoblasts activated by sera under microgravity conditions had a significant 60% reduction in growth (p<0.001) but a paradoxical 2-fold increase in release of the osteoblast autocrine factor PGE2 when compared to ground controls. In addition, a collapse of the osteoblast actin cytoskeleton and loss of focal adhesions has been noted after 4 days in microgravity. Later studies in Biorack on STS-76, 81 and 84 confirmed the increased release of PGE2 and collapse of the actin cytoskeleton in cells grown in microgravity conditions, however flown cells under 1 g conditions maintained normal actin cytoskeleton and fibronectin matrix. The changes seen in the cytoskeleton are probably not due to alterations in fibronectin message or protein synthesis since no differences have been noted in microgravity. Multiple investigators have observed actin and microtubule cytoskeletal modifications in microgravity, suggesting a common root cause for the change in cell architecture. The inability of the O g grown osteoblast to respond to sera activation suggests that there is a major alteration in anabolic signal transduction under microgravity conditions, most probably through the growth factor receptors and/or the associated kinase pathways that are connected to the cytoskeleton. Cell cycle is dependent on the cytoskeleton. Alterations in cytoskeletal structure can block cell growth either in G1 (F-actin microfilament collapse), or in G2/M (inhibition of microtubule polymerization during G2/M-phase). We therefore hypothesize that microgravity would inhibit growth in either G1, or G2/M.