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.     

Performance of a centrifugal phytotron.

It is possible to cultivate plants under an artificial gravity field generated by a centrifugal device in space. In order to determine an optimal magnitude of gravity, there is a need to investigate the relationship between plant and growth and gravity, including not only reduced gravity but also gravity greater than 1G. A prototype centrifugal phytotron was designed and fabricated in order to investigate the relationship between plant growth and increased gravity. This device enables us to cultivate plants over the long term by controlling environmental conditions in the phytotron such as temperature, relative humidity, CO2 concentration and light intensity. The results of our experiment indicate that plant seeds can germinate and grow even under an artificial gravity which changes sinusoidally from 2G to 4G.     

空间飞行与回转器回旋条件下青菜开花与花粉发育的研究

比较研究了SJ-8返回式卫星留轨舱微重力条件与地面三维回转模拟微重力条件下青菜生长与发育情况.研究发现空间微重力条件下青菜开花过程需要大约18 h,明显长于地面对照5 h左右.回转器模拟实验结果表明,改变重力影响了花瓣的伸展与发育及花粉的产量,回转条件下花粉细胞中的微管排列明显不同于静止对照.细胞骨架受到干扰可能是改变重力条件下花粉产量降低的原因之一.本研究首次报道了在空间飞行试验中成功地采用了显微实时图像技术观察植物的开花过程,并获得了从花蕾到开花结束各阶段清晰的图像.      

Living plant systems: How robust are they in the absence of gravity?

The following hierarchical levels can be recognised in plant systems: cells, organs, organisms and gamodemes (interbreeding members of a community). Each level in this ‘living hierarchy’ is both defined and supported by a similar set of sub-systems. Within this framework of plant organization, two complementary questions are relevant for interpreting plant-oriented space experiments: 1) What role, if any, does gravity play in enabling the development of each organizational level? and 2) Does abnormal development in an altered gravity environment indicate sub-system inefficiency? Although a few representatives of the various organizational levels in plant systems have already been the subject of microgravity experiments in space laboratories—from cells in culture to gamodemes, the latter being found in some Closed Environment Life Support Systems—it would be of interest to investigate additional systems with respect to their response to microgravity. Recognition of the sub-systems at each level might be relevant not only for a more complete understanding of plant development but also for the successful cultivation and propagation of plants during long-term space flights and the establishment of plants in extra-terrestrial environments.     

Interaction of gravity with other environmental factors in growth and development: an introduction.

The life of plants and other organisms is governed by the constant force of gravity on earth. The mechanism of graviperception, signal transduction, and gravireaction is one of the major themes in space biology. When gravity controls each step of the life cycle such as growth and development, it does not work alone but operates with the interaction of other environmental factors. In order to understand the role of gravity in regulation of the life cycle, such interactions also should be clarified. Under microgravity conditions in space, various changes are brought about in the process of growth and development. Some changes would be advantageous to organisms, but others would be unfavorable. For overcoming such disadvantages, it may be required to exploit some other environmental factors which substitute for gravity in some properties. In terrestrial plants, gravity can be replaced by light under certain conditions. The gravity-substituting factors may play a principal role in future space development.     

Developing a vitamin greenhouse for the life support system of the International Space Station and for future interplanetary missions.

In order to evaluate the effects of gravity on growing plants, we conducted ground based long-term experiments with dwarf wheat, cultivar Apogee and Chinese cabbage, cultivar Khibinskaya. The test crops had been grown in overhead position with HPS lamp below root module so gravity and light intensity gradients had been in opposite direction. Plants of the control crop grew in normal position under the same lamp. Both crops were grown on porous metallic membranes with stable -1 kPa matric potential on their surface. Results from these and other studies allowed us to examine the differences in growth and development of the plants as well as the root systems in relation to the value of the gravity force influence. Dry weight of the roots from test group was decreased in 2.5 times for wheat and in 6 times - at the Chinese cabbage, but shoot dry biomass was practically same for both test and control versions. A harvest index of the test plants increased substantially. The data shows, that development of the plants was essentially changed in microgravity. The experiments in the space greenhouse Svet aboard the Mir space station proved that it is possible to compensate the effects of weightlessness on higher plants by manipulating gradients of environmental parameters (i.e. photon flux, matric potential in the root zone, etc.). However, the average productivity of Svet concerning salad crops even in ground studies did not provide more than 14 g fresh biomass per day. This does not provide a sufficient level of supplemental nutrients to the crew of the ISS. A cylindrical design of a space plant growth chamber (SPGC) allows for maximal productivity in presence of very tight energy and volume limitations onboard the ISS and provides a number of operational advantages. Productivity from this type of SPGF with a 0.5 kW energy utilization when salad growing would provide approximately 100 g of edible biomass per day, which would almost satisfy requirements for a crew of two in vitamin C and carotene and partly vitamin B group as well as rough fiber.     

Calcium signaling in plant cells in altered gravity.

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

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.     

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.     

Modeling gravity effects on water retention and gas transport characteristics in plant growth substrates

Growing plants to facilitate life in outer space, for example on the International Space Station (ISS) or at planned deep-space human outposts on the Moon or Mars, has received much attention with regard to NASA’s advanced life support system research. With the objective of in situ resource utilization to conserve energy and to limit transport costs, native materials mined on Moon or Mars are of primary interest for plant growth media in a future outpost, while terrestrial porous substrates with optimal growth media characteristics will be useful for onboard plant growth during space missions. Due to limited experimental opportunities and prohibitive costs, liquid and gas behavior in porous substrates under reduced gravity conditions has been less studied and hence remains poorly understood. Based on ground-based measurements, this study examined water retention, oxygen diffusivity and air permeability characteristics of six plant growth substrates for potential applications in space, including two terrestrial analogs for lunar and Martian soils and four particulate substrates widely used in reduced gravity experiments. To simulate reduced gravity water characteristics, the predictions for ground-based measurements (1 − g) were scaled to two reduced gravity conditions, Martian gravity (0.38 − g) and lunar gravity (0.16 − g), following the observations in previous reduced gravity studies. We described the observed gas diffusivity with a recently developed model combined with a new approach that estimates the gas percolation threshold based on the pore size distribution. The model successfully captured measured data for all investigated media and demonstrated the implications of the poorly-understood shift in gas percolation threshold with improved gas percolation in reduced gravity. Finally, using a substrate-structure parameter related to the gaseous phase, we adequately described the air permeability under reduced gravity conditions.     

Ontogeny of plants under various gravity condition.

The results of experiments performed under conditions of microgravity (MG) or under its simulation on the horizontal clinostat (HC) with the callus, seedlings of various species and embryogenic structures have revealed a definite role of gravity as an ecological factor in the processes of cytomorphogenesis, growth, and development. The transformation of differentiated somatic cells of arabidopsis seed into undifferentiated callus was not inhibited under MG, though modifications of the whole callus morphology and of mean cell and nucleus size were observed. The morphogenesis of polar structures such as root-hair bearing cells of Lactuca primary root has been shown to be modified in the course of differentiation under mass acceleration diminished below 0.1 g. Seed germination and seedling morphogenesis under MG follow their normal course, but a significant stimulation of shoot growth with no effect on primary root growth has been determined. A successful in vitro regeneration of Nicotiana tabacum plantlets from leaf cells and subsequent formation of shoots and roots on a continuously rotating HC as well as the formation of viable seeds during seed-to-seed growth of Arabidopsis plants under MG have indicated that gravity plays but a limited role in the processes of embryogenesis and organogenesis.     

Seed-to-seed growth of Arabidopsis thaliana on the International Space Station.

The assembly of the International Space Station (ISS) as a permanent experimental outpost has provided the opportunity for quality plant research in space. To take advantage of this orbital laboratory, engineers and scientists at the Wisconsin Center for Space Automation and Robotics (WCSAR), University of Wisconsin-Madison, developed a plant growth facility capable of supporting plant growth in the microgravity environment. Utilizing this Advanced Astroculture (ADVASC) plant growth facility, an experiment was conducted with the objective to grow Arabidopsis thaliana plants from seed-to-seed on the ISS. Dry Arabidopsis seeds were anchored in the root tray of the ADVASC growth chamber. These seeds were successfully germinated from May 10 until the end of June 2001. Arabidopsis plants grew and completed a full life cycle in microgravity. This experiment demonstrated that ADVASC is capable of providing environment conditions suitable for plant growth and development in microgravity. The normal progression through the life cycle, as well as the postflight morphometric analyses, demonstrate that Arabidopsis thaliana does not require the presence of gravity for growth and development.     

First flight of the ASTROCULTURE (TM) experiment as a part of the U.S. Shuttle/MIR program.

A number of space-based experiments have been conducted to assess the impact of microgravity on plant growth and development. In general, these experiments did not identify any profound impact of microgravity on plant growth and development, though investigations to study seed development have indicated difficulty in plants completing their reproductive cycle. However, it was not clear whether the lack of seed production was due to gravity effects or some other environmental condition prevailing in the unit used for conducting the experiment. The ASTROCULTURE (TM) flight unit contains a totally enclosed plant chamber in which all the critically important environmental conditions are controlled. Normal wheat (Triticum aestivum L.) growth and development in the ASTROCULTURE (TM) flight unit was observed during a ground experiment conducted prior to the space experiment. Subsequent to the ground experiment, the flight unit was transported to MIR by STS-89, as part of the U.S. Shuttle/MIR program, in an attempt to determine if super dwarf wheat plants that were germinated in microgravity would grow normally and produce seeds. The experiment was initiated on-orbit after the flight unit was transferred from the Space Shuttle to MIR. The ASTROCULTURE (TM) flight unit performed nominally for the first 24 hours after the flight unit was activated, and then the unit stopped functioning abruptly. Since it was not possible to return the unit to nominal operation it was decided to terminate the experiment. On return of the flight unit, it was confirmed that the control computer of the ASTROCULTURE (TM) flight unit sustained a radiation hit that affected the control software embedded in the computer. This experience points out that at high orbital inclinations, such as that of MIR and that projected for the International Space Station, the danger of encountering harmful radiation effects are likely unless the electronic components of the flight hardware are resistant to such impacts.     

微重力环境下植物培养箱设计

基于空间微重力下植物的生物学效应及其微重力信号转导研究需要,在微重力条件下培养拟南芥,获得经微重力条件生长的拟南芥样品.在空间实验过程中实时采集、存储和传输植物样品的数字图像,并根据生物样品的生长周期对生物样品进行低温固定和储存,再由返回式卫星带回地面,开展微重力植物生物学效应研究.      

重力对植物细胞生长的效应

从地基研究植物向重性和在神舟八号卫星微重力条件下培养植物细胞出发,探讨重力变化对植物细胞发生作用时产生的生物学效应.已有结果显示,在植物向重性反应和处于失重状态时,重力方向和大小变化对细胞壁代谢具有一定影响.推断细胞形状的维持是由细胞壁的刚性与细胞内膨压平衡所致,当细胞膨压大于细胞壁刚性导致上述平衡打破时就会引起细胞体积增大.因此,重力的变化可能会通过影响植物细胞壁刚性与细胞内膨压的平衡影响细胞生长.      

Progress in plant research in space.

Progress is reviewed of spaceflight research conducted with plants between 1987 and 1992. Numerous plant experiments have been performed on spacecraft and sounding rockets in the past five years by scientists of the US, the former Soviet Union, Europe, and other areas. The experiments are categorized into three areas: gravity sensing, transduction, and response; development and reproduction; and metabolism, photosynthesis, and transport. The results of these experiments continue to demonstrate that gravity and/or other factors of spaceflight affect plants at the organismal, cellular, subcellular, and molecular levels, resulting in changes in orientation, development, metabolism, and growth. The challenge now is to truly dissect the effects of gravity from those of other spaceflight factors and to identify the basic mechanisms underlying gravity's effects.     

Spaceflight hardware for conducting plant growth experiments in space: the early years 1960-2000.

The best strategy for supporting long-duration space missions is believed to be bioregenerative life support systems (BLSS). An integral part of a BLSS is a chamber supporting the growth of higher plants that would provide food, water, and atmosphere regeneration for the human crew. Such a chamber will have to be a complete plant growth system, capable of providing lighting, water, and nutrients to plants in microgravity. Other capabilities include temperature, humidity, and atmospheric gas composition controls. Many spaceflight experiments to date have utilized incomplete growth systems (typically having a hydration system but lacking lighting) to study tropic and metabolic changes in germinating seedlings and young plants. American, European, and Russian scientists have also developed a number of small complete plant growth systems for use in spaceflight research. Currently we are entering a new era of experimentation and hardware development as a result of long-term spaceflight opportunities available on the International Space Station. This is already impacting development of plant growth hardware. To take full advantage of these new opportunities and construct innovative systems, we must understand the results of past spaceflight experiments and the basic capabilities of the diverse plant growth systems that were used to conduct these experiments. The objective of this paper is to describe the most influential pieces of plant growth hardware that have been used for the purpose of conducting scientific experiments during the first 40 years of research.     

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.     

Operations of a spaceflight experiment to investigate plant tropisms

Plants will be an important component in bioregenerative systems for long-term missions to the Moon and Mars. Since gravity is reduced both on the Moon and Mars, studies that identify the basic mechanisms of plant growth and development in altered gravity are required to ensure successful plant production on these space colonization missions. To address these issues, we have developed a project on the International Space Station (ISS) to study the interaction between gravitropism and phototropism in Arabidopsis thaliana. These experiments were termed TROPI (for tropisms) and were performed on the European Modular Cultivation System (EMCS) in 2006. In this paper, we provide an operational summary of TROPI and preliminary results on studies of tropistic curvature of seedlings grown in space. Seed germination in TROPI was lower compared to previous space experiments, and this was likely due to extended storage in hardware for up to 8 months. Video downlinks provided an important quality check on the automated experimental time line that also was monitored with telemetry. Good quality images of seedlings were obtained, but the use of analog video tapes resulted in delays in image processing and analysis procedures. Seedlings that germinated exhibited robust phototropic curvature. Frozen plant samples were returned on three space shuttle missions, and improvements in cold stowage and handing procedures in the second and third missions resulted in quality RNA extracted from the seedlings that was used in subsequent microarray analyses. While the TROPI experiment had technical and logistical difficulties, most of the procedures worked well due to refinement during the project.     

Possible applications of aquatic bioregenerative life support modules for food production in a Martian base.

Water is the essential precondition of life in general and also for the establishment of a Martian base suitable for long duration stays of humans. It is not yet proven if there is indeed a "frozen ocean" under the surface of Mars but if this could be verified it would open innovative aspects for the construction of bioregenerative life support systems (BLSS). In a general concept higher plants will play the predominant role in a Martian BLSS. It is not clear, however, how these will grow and bring seed in reduced gravity and there may be differences in the productivity in comparison to Earth conditions. Therefore, organisms which are already adapted to low gravity conditions, namely non-gravitropic aquatic plants and also aquatic animals may be used to enhance the functionality of the Martian BLSS as a whole. It has been shown already with the so-called C.E.B.A.S. MINIMODULE in the STS-89 and STS-90 spaceshuttle missions that the water plant Ceratophyllum demersum has an undisturbed and high biomass production under space conditions. Moreover, the teleost fish species Xiphophorus helleri adapted easily to the micro-g environment and maintained its normal reproductive functions. Based on this findings a possible scenario is presented in which aquatic plant production modules and combined animal-plant production systems may be used for human food production and water and air regeneration in a Martian base.