Growth restoration in azuki bean and maize seedlings by removal of hypergravity stimuli.

Hypergravity stimuli, gravitational acceleration of more than 1 x g, decrease the growth rate of azuki bean epicotyls and maize coleoptiles and mesocotyls by decreasing the cell wall extensibility via an increase in the molecular mass of matrix polysaccharides. An increase in the pH in the apoplastic fluid is hypothesized to be involved in the processes of the increase in the molecular mass of matrix polysaccharides due to hypergravity. However, whether such physiological changes by hypergravity are induced by normal physiological responses or caused by physiological damages have not been elucidated. In the present study, we examined the effects of the removal of hypergravity stimuli on growth and the cell wall properties of azuki bean and maize seedlings to clarify whether the effects of hypergravity stimuli on growth and the cell wall properties are reversible or irreversible. When the seedlings grown under hypergravity conditions at 300 x g for several hours were transferred to 1 x g conditions, the growth rate of azuki bean epicotyls and maize coleoptiles and mesocotyls greatly increased within a few hours. The recovery of growth rate of these organs was accompanied by an immediate increase in the cell wall extensibility, a decrease in the molecular mass of matrix polysaccharides, and an increase in matrix polysaccharide-degrading activities. The apoplastic pH also decreased promptly upon the removal of hypergravity stimuli. These results suggest that plants regulate the growth rate of shoots reversibly in response to hypergravity stimuli by changing the cell wall properties, by which they adapt themselves to different gravity conditions. This study also revealed that changes in growth and the cell wall properties under hypergravity conditions could be recognized as normal physiological responses of plants. In addition, the results suggest that the effects of microgravity on plant growth and cell wall properties should be reversible and could disappear promptly when plants are transferred from microgravity to 1 x g. Therefore, plant materials should be fixed or frozen on orbit for detecting microgravity-induced changes in physiological parameters after recovering the materials to earth in space experiments.     

Influence of different natural physical fields on biological processes.

In space flight conditions gravity, magnetic, and electrical fields as well as ionizing radiation change both in size, and in direction. This causes disruptions in the conduct of some physical processes, chemical reactions, and metabolism in living organisms. In these conditions organisms of different phylogenetic level change their metabolic reactions undergo changes such as disturbances in ionic exchange both in lower and in higher plants, changes in cell morphology for example, gyrosity in Proteus (Proteus vulgaris), spatial disorientation in coleoptiles of Wheat (Triticum aestivum) and Pea (Pisum sativum) seedlings, mutational changes in Crepis (Crepis capillaris) and Arabidopsis (Arabidopsis thaliana) seedling. It has been found that even in the absence of gravity, gravireceptors determining spatial orientation in higher plants under terrestrial conditions are formed in the course of ontogenesis. Under weightlessness this system does not function and spatial orientation is determined by the light flux gradient or by the action of some other factors. Peculiarities of the formation of the gravireceptor apparatus in higher plants, amphibians, fish, and birds under space flight conditions have been observed. It has been found that the system in which responses were accompanied by phase transition have proven to be gravity-sensitive under microgravity conditions. Such reactions include also the process of photosynthesis which is the main energy production process in plants. In view of the established effects of microgravity and different natural physical fields on biological processes, it has been shown that these processes change due to the absence of initially rigid determination. The established biological effect of physical fields influence on biological processes in organisms is the starting point for elucidating the role of gravity and evolutionary development of various organisms on Earth.     

Growth and development in higher plants under simulated microgravity conditions on a 3-dimensional clinostat.

Growth and development of etiolated pea (Pisum sativum L. cv. Alaska) and maize (Zea mays L. cv. Golden Cross Bantam) seedlings grown under simulated microgravity conditions were intensively studied using a 3-dimensional clinostat as a simulator of weightlessness. Epicotyls of etiolated pea seedlings grown on the clinostat were the most oriented toward the direction far from cotyledons. Mesocotyls of etiolated maize seedlings grew at random and coleoptiles curved slightly during clinostat rotation. Clinostat rotation promoted the emergence of the 3rd internodes in etiolated pea seedlings, while it significantly inhibited the growth of the 1st internodes. In maize seedlings, the growth of coleoptiles was little affected by clinostat rotation, but that of mesocotyls was suppressed, and therefore, the emergence of the leaf out of coleoptile was promoted. Clinostat rotation reduced the osmotic concentration in the 1st internodes of pea seedlings, although it has little effect on the 2nd and the 3rd internodes. Clinostat rotation also reduced the osmotic concentrations in both coleoptiles and mesocotyls of maize seedlings. Cell-wall extensibilities of the 1st and the 3rd internodes of pea seedlings grown on the clinostat were significantly lower and higher as compared with those on 1 g conditions, respectively. Cell-wall extensibility of mesocotyls in seedlings grown on the clinostat also decreased. Changes in cell wall properties seem to be well correlated to the growth of each organ in pea and maize seedlings. These results suggest that the growth and development of plants is controlled under gravity on earth, and that the growth responses of higher plants to microgravity conditions are regulated by both cell-wall mechanical properties and osmotic properties of stem cells.     

Short-term and long-term clinostat and vibration-induced biochemical changes in dwarf marigold stems.

Stems of 21-day dwarf Marigold plants cultivated on the clinostat were compared with plants cultivated on vertical axis rotators ("vibrational controls") and stationary controls for long-term changes in cell wall composition. Stems of 21-day plants grown under stationary conditions and subsequently exposed to the clinostat for 24 hours were also analyzed. Among the long-tern markers, calciun, lignin, and protein-bound hemicellulose (possibly cell wall glycoprotein) clearly differentiated the effects of vibration from those of the clinostat. Short-term differential responses included rate of ethylene production, nastic movement and peroxidase activity of the cell wall, but not of the protoplast.     

The influence of gravity on the structure and functions of plant material

Growth process generate plant form and relate to most physiological functions. The Earth's gravity force affects plant growth in both obvious and subtle ways. It is a major environmental influence on morphology and physiology of plants. Gravity is less important as an agent for plant stress than as an environmental signal to guide growth. The plant's bioaccelerometers are remarkably sensitive, especially in hypogravity. Simulation (clinostat) studies and experiments in satellite laboratories are needed to understand the sensing, transduction, and response characteristics of g related mechanisms. By examining how plants alter growth processes to accomplish developmental or physiological “objectives” we may find it pragmatically desirable to ask ourselves how we might design a plant to achieve such responses to environmental influences. Examples of this design engineering approach for gravity related effects are described as an aid to experimentation.     

地磁扰动期间日本Kokubunji站电离层的扰动特征分析

利用日本Kokubunji站(139.5°E,35.5°N)1959年1月到2004年12月共46年的F2层临界频率foF2参数,统计分析了Kokubunji站电离层F2层峰值电子浓度NmF2随地磁活动、太阳活动、季节和地方时变化的形态特征.结果表明,总体来看,磁暴期间Kokubunji站电离层响应以正暴为主,其中在太阳高年夏季为负暴,冬季为正暴,春秋季以负暴为主但幅度较小;在太阳低年夏季以正暴为主,冬季为正暴,春秋季以正暴为主.NmF2扰动与ap指数在夏季太阳高年负相关,在冬季无论太阳高年低年均为正相关,春秋季中4月和9月在太阳高年类似夏季,3月和10月在太阳低年类似冬季.电离层最大负相扰动对最大地磁活动的延迟时间约为12～15 h;正相扰动的延迟时间则分别为3 h和10 h.地磁活跃期间地方时黄昏后到午夜前倾向于正相扰动,清晨倾向于负相扰动.      

Mechanosensing and signal transduction in tendrils.

The perception of thigmic stimuli is a widespread phenomenon among plants with decisive meaning for the ability to survive. Beside a general sensitivity for mechanical stimuli many plants have evolved specialized organs with highly developed mechanisms to perceive and transduce the applied forces. Tendrils of Bryonia dioica and Pisum sativum have been chosen to study the effects of mechanical stimulation on plant physiology. Both types of tendrils, although exhibiting different morphology, respond to such a stimulus with a rapid coiling response to the dorsal side of the organ within minutes. The actual perception of the stimulus is most likely coupled to the cytoskeleton serving as the mediator between the physical stimulus and the biochemical response. Drugs affecting the status of the cytoskeleton were used to get more insights into this specific process. The results indicate that microtubuli (MT) play the most important role in the perception of thigmic stimuli in tendrils. Colchicine-mediated disruption of MT lead to total inhibition of the response to the thigmic stimulus in tendrils of Pisum and to a reduced response in Bryonia. Alamethicin, an ionophore that can mimic action potentials in membranes, was able to bypass this inhibition suggesting a direct involvement of MT in depolarization of the membranes. Auxin, however, which is also supposed to be involved in the regulation of the coiling response, failed to bypass colchicine-dependent inhibition. Vinblastine, another microtubule depolimerizing agent, did induce tendril coiling in Pisum without further stimulation. Application of taxol and other MT-stabilizing drugs as well as disruption of the actin network did not affect the coiling response of tendrils. In Pisum indole-3-acetic acid (IAA) is induced after mechanical stimulation during the coiling response, but not jasmonic acid. A further consequence of mechanical stimulation is the induction of an oxidative burst and an increase in soluble sugar. A model is presented integrating these results and might serve as a common basis for the understanding of the perception of mechanical stimuli.     

Evaluation of experiments involving the study of plant orientation and growth under different gravitational conditions.

The manifestation of gravitropic reaction in plants has been considered from the phylogenetic point of view. A chart has been suggested according to which it is supposed that the first indications of the ability to identify the direction of the gravitational vector were inherent in the most ancient eukaryotes, which gave rise to green, brown, yellow-green, golden and diatomaceous algae as well as fungi. The experiments on the role of gravity in plant ontogenesis are being continued. The sum total of the data obtained in a number of experiments in space shows that under these conditions a structurally modified but normally functioning gravireceptive apparatus is formed. The data confirming the modification, under changed gravity, of the processes of integral and cellullar growth of the axial organs of seedlings as well as of the anatomo-morphological structure and developmental rates of plants during their prolonged growth in space are presented. It is assumed that this fact testifies to the presence of systems interacting with gravity during plant ontogenesis. At the same time the necessity for further experiments in order to differentiate an immediate biological effect of gravity from the ones conditioned by it indirectly due to the changes in the behavior of liquids and gases is pointed out. The methodological aspects of biological experiments in space as the main source of reliable information on the biological role of gravity are discussed.     

Robust H∞ controller design for attitude stabilization of flexible spacecraft with input constraints

This paper addresses the attitude stabilization and vibration suppression problem for flexible spacecraft subject to model parameter uncertainty, controller perturbations, external disturbances and input constraints. The attitude model of flexible spacecraft is described and converted into a state space form in terms of passive and active vibration suppression schemes. A novel state feedback controller is proposed based on the exactly available expectation of a new variable, which is introduced to model a randomly occurring controller gain perturbation. Based on Lyapunov stability theory, sufficient conditions for the existence of the nonfragile H_∞ controller considering input constraints are given based on linear matrix inequalities (LMIs) in terms of additive perturbation and multiplicative perturbation. Then, the developed controller subject to required constraints can be obtained, where the nonfragile property is fully considered to improve the tolerance to uncertainties in the controller. Numerical simulations are performed to demonstrate the effectiveness and superiority of the proposed control strategy in attitude stabilization and vibration suppression, where it should be noted that the passive vibration suppression scheme is superior for high natural frequencies while the active vibration suppression scheme is superior for low natural frequencies. Moreover, the low natural frequencies have more influence on the performance of attitude stabilization and vibration suppression.     

The effect of radiation on the long term productivity of a plant based CELSS.

Mutations occur at a higher rate in space than under terrestrial conditions, primarily due to an increase in radiation levels. These mutations may effect the productivity of plants found in a controlled ecological life support system (CELSS). Computer simulations of plants with different ploidies, modes of reproduction, lethality thresholds, viability thresholds and susceptibilities to radiation induced mutations were performed under space normal and solar flare conditions. These simulations identified plant characteristics that would enable plants to retain high productivities over time in a CELSS.     

Changes in hyperspectral reflectance signatures of lettuce leaves in response to macronutrient deficiencies

The adaptation of specific remote sensing and hyperspectral analysis techniques for the determination of incipient nutrient stress in plants could allow early detection and precision supplementation for remediation, important considerations for minimizing mass of advanced life support systems on space station and long term missions. This experiment was conducted to determine if hyperspectral reflectance could be used to detect nutrient stress in Lactuca sativa L. cv. Black Seeded Simpson. Lettuce seedlings were grown for 90 days in a greenhouse or growth chamber in vermiculite containing modified Hoagland’s nutrient solution with key macronutrient elements removed in order to induce a range of nutrient stresses, including nitrogen, phosphorus, potassium, calcium, and magnesium. Leaf tissue nutrient concentrations were compared with corresponding spectral reflectances taken at the end of 90 days. Spectral reflectances varied with growing location, position on the leaf, and nutrient deficiency treatment. Spectral responses of lettuce leaves under macronutrient deficiency conditions showed an increase in reflectance in the red, near red, and infrared wavelength ranges. The data obtained suggest that spectral reflectance shows the potential as a diagnostic tool in predicting nutrient deficiencies in general. Overlapping of spectral signatures makes the use of wavelengths of narrow bandwidths or individual bands for the discrimination of specific nutrient stresses difficult without further data processing.     

Dust Acoustic Wave in Dusty Plasmas With Streaming Ions Under Ultraviolet Irradiation

The reductive perturbation method is applied to investigate the dust acoustic soliton in dusty plasmas with streaming ions under ultraviolet irradiation theoretically and numerically. The self-consistent dust charge variation is taken into account. It is shown that the ultraviolet irradiation can significantly lower the magnitude of the dust negative charge, and ion streaming velocity firstly raise the magnitude of the dust negative charge and then lower it. With the growth of (Ultraviolet) UV photo flux or ion streaming velocity, the phase velocity and width of the solitary waves decrease, whereas its amplitude increases.      

Growth of pea epicotyl in low magnetic field implication for space research

A magnetic field is an inescapable environmental factor for plants on the earth. However, its impact on plant growth is not well understood. In order to survey how magnetic fields affect plant, Alaska pea seedlings were incubated under low magnetic field (LMF) and also in the normal geo-magnetic environment. Two-day-old etiolated seedlings were incubated in a magnetic shield box and in a control box. Sedimentation of amyloplasts was examined in the epicotyls of seedlings grown under these two conditions. The elongation of epicotyls was promoted by LMF. Elongation was most prominent in the middle part of the epicotyls. Cell elongation and increased osmotic pressure of cell sap were found in the epidermal cells exposed to LMF. When the gravitational environment was 1G, the epicotyls incubated under both LMF and normal geomagnetic field grew straight upward and amyloplasts sedimented similarly. However, under simulated microgravity (clinostat), epicotyl and cell elongation was promoted. Furthermore, the epicotyls bent and amyloplasts were dispersed in the cells in simulated microgravity. The dispersion of amyloplasts may relate to the posture control in epicotyl growth under simulated microgravity generated by 3D clinorotation, since it was not observed under LMF in 1G. Since enhanced elongation of cells was commonly seen both at LMF and in simulated microgravity, all elongation on the 3D-clinostat could result from pseudo-low magnetic field, as a by-product of clinorotation. (i.e., clinostat results could be based on randomization of magnetic field together with randomization of gravity vector.) Our results point to the possible use of space for studies in magnetic biology. With space experiments, the effects of dominant environmental factors, such as gravity on plants, could be neutralized or controlled for to reveal magnetic effects more clearly.     

Influence of nonuniform magnetic fields on orientation of plant seedlings in microgravity conditions.

Experiments on the spatial behavior of the flax (Linum usitatissimum, L.) seedlings in a nonuniform magnetic field were conducted on the orbital space stations "Salut" and "Mir". This field can displace sensory organelles (statoliths) inside receptor cells and such displacement should cause a physiological reaction of the plant-tropistic curvature. Experiments were conducted in the custom-built "Magnetogravistat" facility, where seeds were germinated and grown for 3-4 days in a magnetic field with the dynamic factor grad (H2/2) approximately equal to 10(7) Oe2/cm, then fixed on orbit and returned to Earth for analysis. It was found, that 93% of the seedlings were oriented in the field consistently with curvature in response to displacement of statoliths along the field gradient by ponderomotive magnetic forces, while control seedlings grew in the direction of the initial orientation of the seed. This suggests, that gravity receptors of plants recognized magnetic forces on statoliths as gravity, and that gravity stimulus can be substituted for plants by a force of a different physical nature.     

Effect of salt stress on growth and physiology in amaranth and lettuce: Implications for bioregenerative life support system

Growing plants can be used to clean waste water in bioregenerative life support system (BLSS). However, NaCl contained in the human urine always restricts plant growth and further reduces the degree of mass cycle closure of the system (i.e. salt stress). This work determined the effect of NaCl stress on physiological characteristics of plants for the life support system. Amaranth (Amaranthus tricolor L. var. Huahong) and leaf lettuce (Lactuca sativa L. var. Luoma) were cultivated at nutrient solutions with different NaCl contents (0, 1000, 5000 and 10,000 ppm, respectively) for 10 to 18 days after planted in the Controlled Ecological Life Support System Experimental Facility in China. Results showed that the two plants have different responses to the salt stress. The amaranth showed higher salt-tolerance with NaCl stress. If NaCl content in the solution is below 5000 ppm, the salt stress effect is insignificant on above-ground biomass output, leaf photosynthesis rate, Fv/Fm, photosynthesis pigment contents, activities of antioxidant enzymes, and inducing lipid peroxidation. On the other hand, the lettuce is sensitive to NaCl which significantly decreases those indices of growth and physiology. Notably, the lettuce remains high productivity of edible biomass in low NaCl stress, although its salt-tolerant limitation is lower than amaranth. Therefore, we recommended that amaranth could be cultivated under a higher NaCl stress condition (<5000 ppm) for NaCl recycle while lettuce should be under a lower NaCl stress (<1000 ppm) for water cleaning in future BLSS.     

The mechanism by which an asymmetric distribution of plant growth hormone is attained.

Zea mays (sweet corn) seedlings attain an asymmetric distribution of the growth hormone indole-3-acetic acid (IAA) within 3 minutes following a gravity stimulus. Both free and esterified IAA (that is total IAA) accumulate to a greater extent in the lower half of the mesocotyl cortex of a horizontally placed seedling than in the upper half. Thus, changes in the ratio of free IAA to ester IAA cannot account for the asymmetric distribution. Our studies demonstrate there is no de novo synthesis of IAA in young seedlings. We conclude that asymmetric IAA distribution is attained by a gravity-induced, potential-regulated gating of the movement of IAA from kernel to shoot and from stele to cortex. As a working theory, which we call the Potential Gating Theory, we propose that perturbation of the plant's bioelectric field, induced by gravity, causes opening and closing of transport channels in the plasmodesmata connecting the vascular stele to the surrounding cortical tissues. This results in asymmetric growth hormone distribution which results in the asymmetric growth characteristics of the gravitropic response.     

Spectral composition of light and plant productivity.

Among other problems the Institute of Biophysics is working on the development of physiological and fundamental aspects of intensive light cultivation of higher plants. These technologies can be used in life support systems for stationary space station such as a Lunar base, a planetary base or a large orbital station. The source of energy may be the Sun or a nuclear reactor. In certain conditions, such sources of energy allow the use of a very broad range of irradiance of plants, in particular in the light energy range up to 2-3 times the solar energy (up to 100-1200 W/m2 PAR). Our Institute was the first to show that under such a high irradiance, some plants (radish, wheat, for example) can actively photosynthesize and exhibit high productivity on a sowing area basis. These results were later confirmed in the laboratory of Prof. Salisbury (USA).     

Effects of air velocity on photosynthesis of plant canopies under elevated CO2 levels in a plant culture system.

To obtain basic data for adequate air circulation for promoting plant growth in closed plant production modules in bioregenerative life support systems in space, effects of air velocities ranging from 0.1 to 0.8 m s-1 on photosynthesis in tomato seedlings canopies were investigated under atmospheric CO2 concentrations of 0.4 and 0.8 mmol mol-1. The canopy of tomato seedlings on a plug tray (0.4 x 0.4 m2) was set in a wind-tunnel-type chamber (0.6 x 0.4 x 0.3 m3) installed in a semi-closed-type assimilation chamber (0.9 x 0.5 x 0.4 m3). The net photosynthetic rate in the plant canopy was determined with the differences in CO2 concentrations between the inlet and outlet of the assimilation chamber multiplied by the volumetric air exchange rate of the chamber. Photosynthetic photon flux (PPF) on the plant canopy was kept at 0.25 mmol m-2 s-1, air temperature at 23 degrees C and relative humidity at 55%. The leaf area indices (LAIs) of the plant canopies were 0.6-2.5 and plant heights were 0.05-0.2 m. The net photosynthetic rate of the plant canopy increased with increasing air velocities inside plant canopies and saturated at 0.2 m s-1. The net photosynthetic rate at the air velocity of 0.4 m s-1 was 1.3 times that at 0.1 m s-1 under CO2 concentrations of 0.4 and 0.8 mmol mol-1. The net photosynthetic rate under CO2 concentrations of 0.8 mmol mol-1 was 1.2 times that under 0.4 mmol mol-1 at the air velocity ranging from 0.1 to 0.8 m s-1. The results confirmed the importance of controlling air movement for enhancing the canopy photosynthesis under an elevated CO2 level as well as under a normal CO2 level in the closed plant production modules.     

Induction of hypoxic root metabolism results from physical limitations in O2 bioavailability in microgravity.

Numerous spaceflight experiments have noted changes in the roots that are consistent with hypoxia in the root zone. These observations include general ultrastructure analysis and biochemical measurements to direct measurements of stress specific enzymes. In experiments that have monitored alcohol dehydrogenase (ADH), the data shows this hypoxically responsive gene is induced and is associated with increased ADH activity in microgravity. These changes in ADH could be induced either by spaceflight hypoxia resulting from inhibition of gravity mediated O2 transport, or by a non-specific stress response due to inhibition of gravisensing. We tested these hypotheses in a series of two experiments. The objective of the first experiment was to determine if physical changes in gravity-mediated O2 transport can be directly measured, while the second series of experiments tested whether disruption of gravisensing can induce a non-specific ADH response. To directly measure O2 bioavailability as a function of gravity, we designed a sensor that mimics metabolic oxygen consumption in the rhizosphere. Because of these criteria, the sensor is sensitive to any changes in root O2 bioavailability that may occur in microgravity. In a KC-135 experiment, the sensor was implanted in a moist granular clay media and exposed to microgravity during parabolic flight. The resulting data indicated that root O2 bioavailability decreased in phase with gravity. In experiments that tested for non-specific induction of ADH, we compared the response of transgenic Arabidopsis plants (ADH promoted GUS marker gene) exposed to clinostat, control, and waterlogged conditions. The plants were grown on agar slats in a growth chamber before being exposed to the experimental treatments. The plants were stained for GUS activity localization, and subjected to biochemical tests for ADH, and GUS enzyme activity. These tests showed that the waterlogging treatment induced significant increases in GUS and ADH enzyme activities, while the control and clinostat treatments showed no response. This work demonstrates: (1) the inhibition of gravity-driven convective transport can reduce the O2 bioavailability to the root tip, and (2) the perturbation of gravisensing by clinostat rotation does not induce a nonspecific stress response involving ADH. Together these experiments support the microgravity convection inhibition model for explaining changes in root metabolism during spaceflight.     

Somatic embryos of daylily in space

Poor growth and nuclear abnormalities observable in some space-grown plants have been hypothesized as due to a combination of factors such as degree of development, the specific way the plants are grown and the way they experience multiple stresses, some of which are space-specific. Data from a 132-day experiment on ‘Mir’ using embryogenic cell cultures of daylily (Hemerocallis) allow seemingly contradictory evidence from earlier Shuttle missions to be harmonized: a) the more developed an embryo the less likely it is to suffer catastrophic cell stress during growth, whereas the less developed it is, the greater its vulnerability; (b) the extent to which the stress becomes manifest is also dependent on the extent of pre-existing stresses imposed by suboptimal growing conditions; (c) an appropriate, albeit undesirable, ‘stress match’ with other non-equilibrium determinants, much like a ‘tug of war’, can result in genomic variations in space. It is not understood what is/are the feature(s) of the space environment that cause the various cell division perturbations but they have not yet been mimicked on earth. The stress symptoms were found only in space materials and, as predicted, they were most frequently encountered in smaller, less-developed materials grown under non-optimized conditions. It is concluded that, while any substantial deviation from ‘optimum’ can be a ‘stress’, spaceflight subjects vulnerable materials to cell division or DNA-repair stress(es) that appear distinctive, but remain elusive so far. Fastidiously-controlled growing environments must be devised to resolve the matter of direct versus indirect effects of space. On a practical level, it is predicted that adapting plant biotechnologies to space conditions will not be a casual matter.