Plant growth, development and embryogenesis during Salyut-7 flight.

The growth and geotropic movements of roots and hypocotyls of lettuce have been studied on board the Salyut 7 station in a stationary position and on the centrifuge at 0.01, 0.1 and 1 g. On the centrifuge at 0.1 and 0.01 g as well as under weightlessness, the final length of hypocotyls was by 8-16% greater than in control plants on the centrifuge at 1 g. The length of roots, however, was reduced by 17% at 0.01 g and under weightlessness; at 0.1 g their growth is much the same as at 1 g. On the Earth, while growing in a vertical position, and in space at 0 < or = g, the roots and hypocotyls deviate from the longitudinal axis of the seed. Average values of deviation eagles on the Earth are always equal to zero, while this is not always the case in space, which indicates the biological effect of microgravity conditions on board a spacecraft. The threshold of geotropic sensitiveness of lettuce hypocotyls, calculated from the linear regression parameters of the dependence of the response geotropic reaction upon the value of the centrifugal force, comprised 2.9 x 10(-3) g. In the Fiton 3 micro-greenhouse under spaceflight conditions, the plants of Arabidopsis thaliana (L) Heynh have, for the first time, undergone a full cycle of individual development. The seeds sown during the flight germinated, performed growth processes, formed vegetative and generative organs and, judging by the final result, they succeeded in fecundation, embryogenesis and ripening. Despite the noted modification of growth and development of plants in space, 42% of formed seeds appeared to be valuable biologically.     

The state of gravity sensors and peculiarities of plant growth during different gravitational loads.

In the primary roots of lettuce shoots grown under altered gravitational conditions--180 degrees inversion on the centrifuged clinostat, horizontal clinostat and in dynamic weightlessness--localization of the cellular organelles, cell morphology and peculiarities of growth have been studied. Significant changes took place in the localization of amyloplasts on the horizontal clinostat. The changes of amyloplast position in the cap cells on the horizontal clinostat and under weightlessness are similar. A change of the normal shoot position (180 degrees inversion and horizontal clinostat) causes an inhibition of growth. Weightlessness increases the length of axial organs and cells in the zone of elongation, but decreases the nitotic index in comparison to the centrifuged control. The anlysis of the formation of generative organs has been carried out for Arabidopsis plants grown on board the orbital station Salyut-6. The ability of plants to undergo vegetative growth and to pass through early phases of generative development under weightlessness was confirmed.     

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.     

Biological effects of weightlessness and clinostatic conditions registered in cells of root meristem and cap of higher plants.

Research in cellular reproduction, differentiation and vital activity, i.e. processes underlying the development and functioning of organisms, plants included, is essential for solving fundamental and applied problems of space biology. Detailed anatomical analysis of roots of higher plants grown on board the Salyut 6 orbital research station show that under conditions of weightlessness for defined duration mitosis, cytokinesis and tissue differentiation in plant vegetative organs occur essentially normally. At the same time, certain rearrangements in the structural organization of cellular organelles--mainly the plastid apparatus, mitochondria, Golgi apparatus and nucleus--are established in the root meristem and cap of the experimental plants. This is evidence for considerable changes in cellular metabolism. The structural changes in the subcellular level arising under spaceflight conditions are partially absent in clinostat experiments designed to simulate weightlessness. Various clinostatic conditions have different influences on the cell structural and functional organization than does space flight. It is suggested that alterations of cellular metabolism under weightlessness and clinostatic conditions occur within existing genetic programs.     

Increase of atmospheric CO2: response patterns of a simple terrestrial man-made ecosystem.

Simple models of terrestrial ecosystems with a limited number of components are an efficient tool to study the main laws of functioning of populations, including microbial ones, and their communities, as components of natural ecosystems, under variable environmental conditions. Among other factors are the increase of carbon dioxide in the atmosphere and limitation of plants' growth by biogenic elements. The main types of ecosystems' responses to changes in environmental conditions (a change in CO2 concentration) have been demonstrated in a "plants-rhizospheric microorganisms-artificial soil" simple experimental system. The mathematical model of interactions between plants and microorganisms under normal and elevated atmospheric CO2 and limitation by nutrients (nitrogen and phosphorus) yielded a qualitative agreement between calculated and experimental values of limiting substances concentrations and release rates of exudates.     

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.     

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.     

Plant reproduction systems in microgravity: experimental data and hypotheses.

Elucidation of the possibilities for higher plants to realize complete ontogenesis, from seed to seed, and to propagate by seeds in microgravity, is a fundamental task of space biology connected with the working of the CELSS program. At present, there are results of only 6 spaceflight experiments with Arabidopsis thaliana, an ephemeral plant which will flower and fruit in orbit. Morphogenesis of generative organs occurs normally in microgravity, but unlike the ground control, buds and flowers mainly contain sterile elements of the androecium and gynoecium which degenerate at different stages of development in microgravity. Cytological peculiarities of male and female sterility in microgravity are similar to those occurring naturally during sexual differentiation. Many of the seed formed in microgravity do not contain embryos. Hypotheses to explain abnormal reproductive development in microgravity are: 1) nutritional deficiency, 2) insufficient light, 3) intensification of the influence of the above-mentioned factors by microgravity, 4) disturbances of a hormonal nature, and 5) the absence of pollination and fertilization. Possible ways for testing these hypotheses and obtaining viable seeds in microgravity are discussed.     

Capillary movement of liquid in granular beds in microgravity.

A more complete understanding of the dynamics of capillary flow through an unsaturated porous medium would be useful for the development of an effective water and nutrient delivery system for the growth of plants in space. An experiment was conducted on the Mir Space Station that used an experimental cuvette called "Capillary Test Bed" to compare fluid migration under terrestrial laboratory conditions by positioning the cuvette such that the hydrostatic force is negated and on Mir under microgravity conditions. Differences in fluid migration in the cuvette were observed with migration being slower in microgravity compared with some ground control experiments.     

Possible use of a 3-D clinostat to analyze plant growth processes under microgravity conditions.

A three-dimensional (3-D) clinostat equipped with two rotation axes placed at right angles was constructed, and various growth processes of higher plants grown on this clinostat were compared with ground controls, with plants grown on the conventional horizontal clinostat, and with those under real microgravity in space. On the 3-D clinostat, cress roots developed a normal root cap and the statocytes showed the typical polar organization except a random distribution of statoliths. The structural features of clinostatted statocytes were fundamentally similar to those observed under real microgravity. The graviresponse of cress roots grown on the 3-D clinostat was the same as the control roots. On the 3-D clinostat, shoots and roots exhibited a spontaneous curvature as well as an altered growth direction. Such an automorphogenesis was sometimes exaggerated when plants were subjected to the horizontal rotation, whereas the curvature was suppressed on the vertical rotation. These discrepancies in curvature between the 3-D clinostat and the conventional ones appear to be brought about by the centrifugal force produced. Thus, the 3-D clinostat was proven as a useful device to simulate microgravity.     

Pea chloroplasts under clino-rotation: lipid peroxidation and superoxide dismutase activity.

The lipid peroxidation (LP) intensity and the activity of the antioxidant enzyme superoxide dismutase (SOD) were studied in chloroplasts of pea (Pisum sativum L.) plants grown for 7 and 14 days under clino-rotation. An increase in LP levels in chloroplasts during both terms of clinorotation in comparison with stationary controls was documented. SOD activity increased in chloroplasts of plants that were clino-rotated for seven days. SOD has a significant protective effect by diminishing the availability of O2-. However, under more prolonged clino-rotation (14 days), SOD activity decreased but was still higher than in the control samples. In accordance with Selye's oxidative stress theory (Selye, 1956; modified by Leshem et al., 1998), plants that were clino-rotated for seven days are presumed to be in a stage of resistance while 14-day plants reached a stage of exhaustion.     

Short-term and long-term effects of low total pressure on gas exchange rates of spinach.

In this study, spinach plants were grown under atmospheric and low pressure conditions with constant O2 and CO2 partial pressures, and the effects of low total pressure on gas exchange rates were investigated. CO2 assimilation and transpiration rates of spinach grown under atmospheric pressure increased after short-term exposure to low total pressure due to the enhancement of leaf conductance. However, gas exchange rates of plants grown at 25 kPa total pressure were not greater than those grown at atmospheric pressure. Stomatal pore length and width were significantly smaller in leaves grown at low total pressure. This result suggested that gas exchange rates of plants grown under low total pressure were not stimulated even with the enhancement of gas diffusion because the stomatal size and stomatal aperture decreased.     

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.     

Ultrastructural and growth indices of Chlorella culture in multicomponent aquatic systems under space flight conditions.

Submicroscopic organization of Chlorella cells cultivated under space flight conditions in three-component aquatic system has been studied. Comparison of the experimental cells with that of the controls revealed certain rearrangements of cell organelles particularly, a reduction in the amount of reserve polysaccharides in chloroplasts, increase of cell vacuolization and mitochondrion volume, complication configuration of plasmalemma evaginations and invaginations, and also disturbances in the process of cytokinesis. More over an increase in the number of Chlorella cells infected by bacteria was shown in the experimental variant. No considerable differences were established in the growth characteristics of the experimental and control populations. A comparative cytological analysis revealed general regularities of organelles in Chlorella cells cultivated under space flight condition in the uni- and multicomponent systems.     

The effect of microgravity on the development of plant protoplasts flown on Biokosmos 9.

An experiment using plant protoplasts has been accepted for the IML-1 Space Shuttle mission scheduled for 1991. Preparatory experiments have been performed using both fast and slow rotating clinostats and in orbit to study the effect of simulated and real weightlessness on protoplast regeneration. Late access to the space vehicles before launch has required special attention since it is important to delay cell wall regeneration until the samples are in orbit. On a flight on Biokosmos 9 ("Kosmos-2044") in September 1989 some preliminary results were obtained. Compared to the ground control, the growth of both carrot and rapeseed protoplasts was decreased by 18% and 44% respectively, after 14 days in orbit. The results also indicated that there is less cell wall regeneration under micro-g conditions. Compared to the ground controls the production of cellulose in rapeseed and carrot flight samples was only 46% and 29% respectively. The production of hemicellulose in the flight samples was 63% and 67% respectively of that of the ground controls. In both cases all samples reached the stage of callus development. The peroxidase activity was also found to be lower in the flight samples than in the ground controls, and the number of different isoenzymes was decreased in the flight samples. In general, the regeneration processes were retarded in the flight samples with respect to the ground controls. From a simulation experiment for IML-1 performed in January 1990 at ESTEC, Holland, regenerated plants have been obtained. These results are discussed and compared to the results obtained on Biokosmos 9. Protoplast regeneration did not develop beyond the callus stage in either the flight or the ground control samples from the Biokosmos 9 experiment.     

Spectral quality may be used to alter plant disease development in CELSS.

Plants were grown under light emitting diode (LED) arrays with different spectral qualities to determine the effects of light on the development of tomato mosaic virus (ToMV) in peppers and powdery mildew on cucumbers. One LED array supplied 100% of the photosynthetic photon flux (PPF) at 660 nm, a second array supplied 90% of the PPF at 660 nm and 10% at 735 nm, and a third array supplied 98% of the PPF at 660 nm with 2% in the blue region (380-500 nm) supplied by blue fluorescent lamps. Control plants were grown under metal halide (MH) lamps. Pepper plants inoculated with ToMV and grown under 660 and 660/735 LED arrays showed marked increases in both the rate and the severity of symptoms as compared to inoculated plants grown under the MH lamp or 660/blue array. Pepper plants grown under the 660/blue array did not develop symptoms as rapidly as inoculated plants grown under the 660 or 660/735 arrays, but they did develop symptoms faster than inoculated plants grown under the MH lamp. The numbers of colonies of powdery mildew per leaf and the size of each colony were greatest on inoculated cucumber plants grown under the MH lamp.     

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.     

Gravity as an obligatory factor in normal higher plant growth and development

Gravity may influence different aspects of plant activity. The present report deals with two questions: gravity as an ecological factor determining spatial orientation of plant growth; and second, a possible requirement for gravity in the process of normal growth, morphogenesis and generative development of plants.     

Clinostating effects on biochemical characteristics and productivity of healthy and virus-infected wheat plants of dwarf Apogee variety.

The effects of clinostating on physiological processes and biochemical characteristics of wheat plants (Triticum aestivum L.) both healthy and infected by the wheat streak mosaic virus (WSMV) were studied. In six experiments, each lasting over 30 days, healthy and infected plants of the dwarf Apogee variety were grown under conditions of continuous horizontal and vertical clinostating with 2 rpm at 21 +/- 2 degrees C and 6000 1x (the optimal moisture of a substrate being maintained). The control variants (healthy and infected) were simultaneously grown under the same conditions of temperature and illumination in stationary containers and in open pots. During the experiment, visual observations were carried out over the state of tested plants. After completing the experiment, biometric indices, pigment, carbohydrate and dry matter contents were determined in all the plants. It was shown that clinostating sharply reduced the reproductive function of healthy plants and considerably affected their biomass (productivity) and concentration of chlorophylls and sugars. The viral infection resulted in further reduction of these characteristics. In control variants the viral effect was more significant. We speculate that clinostating reduced the rate of reproduction and spread of the virus.     

Gravity effects on connective tissue biosynthesis by cultured mesenchymal cells.

Quantitative and qualitative aspects of collagen synthesis under microgravity, normal gravity and hypergravity conditions were investigated during the spacelab D-2 mission by incubating human fibroblast cultures with [3H]-proline for 0, 4, 7, 10 and 20 hours. Quantitative analysis revealed an increase of collagen synthesis under microgravity conditions, being 40% higher than 1g controls. Hypergravity samples at 1.44g, 6.6g and 10g showed a decrease in collagen synthesis with increasing g, being down to about 15% at 10g. The relative proportion of collagen from total protein synthesized, the secretion of collagen by the cells, proline hydroxylation of individual collagen alpha-chains and the relative proportions of collagens I, III and V synthesized were not affected at any of the applied conditions.