Force sensitivity of plant gravisensing.

Rotation at 4, 10, 50 and 100 rpm on a horizontal clinostat and in microgravity exerts limited effects on the morphogenesis of lettuce and cress root statocytes and statoliths if compared with the vertical control or 1 g spaceflight reference centrifuge. However, the average distance of statoliths from the distal wall increases. The pattern of plastid location of microgravity-grown and that of clino-rotated samples has been determined at 10, 50, and 100 rpm. Experiments on the centrifuge-clinostat and spaceflight centrifuge (acceleration forces of 0.005 to 1 g) revealed that the average statolith location depends on the amplitude of acropetally or basipetally directed mass acceleration. Decreasing the acropetally directed force from 1 g to 0.4 g dislocates statoliths towards the cell center possibly mediated by the elastic forces of the cytoskeleton. In statocytes formed on the clinostat or in microgravity, the majority of statoliths are located at the center of the cell. To force the statoliths from the center of the statocyte towards one of its poles, a threshold mass acceleration of 0.01 g is required. Statocytes with centrally-located statoliths are considerably more effective in transducing a gravistimulus than those with distally-located plastids. The latent time of the graviresponse is shorter and the response itself is enhanced in roots grown on the clinostat compared to vertically grown samples. The early phases of graviperception are independent of root growth conditions since presentation time and g-threshold are similar for roots grown stationary and those on a clinostat. We propose a sequence of events in gravitropic stimulation that considers not only the lateral displacement of statoliths, as predicted by the starch-statolith hypothesis, but also its longitudinal motion, together with differential gravisensitivity of mechanotransducing structures along the lower-most longitudinal cell wall.     

The effects of HGMFs on the plant gravisensing system.

High Gradient Magnetic Fields (HGMFs) offer new opportunities for studying the gravitropic system of plants. However, it is necessary to analyze the influence that HGMF can have on cellular processes and structures that may not be related to amyloplasts displacement. This paper considers possible HGMF effects on plants, which may accompany HGMF stimulation of amyloplasts and contribute to the mechanisms of the HGMF-induced curvature.     

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.     

Gravitropic bending of fruiting bodies--a model based on hyphal gravisensing and cooperativity.

Gravitropic bending of the winter mushroom Flammulina velutipes is achieved by differential growth of the apical part of the stem, the transition zone. Ultrastructural analysis revealed that bending is due to the relaxation of tissue tensions at the lower flank of the stem where hyphal extension growth is promoted in contrast to the upper flank. Extension of lower flank hyphae is preceded by a conspicuous accumulation of microvesicles in the cytosol and their subsequent fusion with the vacuolar compartment, leading to a large volume increase. The hypothesis is put forward that all hyphae in the transition zone are capable of gravisensing. It is derived from experiments with transition zone segments, which exhibit negative gravitropic response independent from their origin within the stem. A model is presented which connects individual gravisensing of the hyphae with a cooperative response within the stem or small segments of the stem. An essential step is the transmission of positional information, by each hypha with respect to the gravitational vector, to the surroundings. The existence of a soluble growth regulator, which is enriched at the lower flank of the stem, is discussed. A gradient could be formed which precedes the gradient of microvesicle formation, and thereby determines the change of growth direction.     

Theoretical and practical considerations for staggered production of crops in a BLSS.

A functional Bioregenerative Life Support System (BLSS) will generate oxygen, remove excess carbon dioxide, purify water, and produce food on a continuous basis for long periods of operation. In order to minimize fluctuations in gas exchange, water purification, and yield that are inherent in batch systems, staggered planting and harvesting of the crop is desirable. A 418-d test of staggered production of potato cv. Norland (26-d harvest cycles) using nutrients recovered from inedible biomass was recently completed at Kennedy Space Center. The results indicate that staggered production can be sustained without detrimental effects on life support functions in a CELSS. System yields of H2O, O2 and food were higher in staggered than batch plantings. Plants growing in staggered production or batch production on "aged" solution initiated tubers earlier, and were shorter than plants grown on "fresh" solution. This morphological response required an increase in planting density to maintain full canopy coverage. Plants grown in staggered production used available light more efficiently than the batch planting due to increased sidelighting.     

Oscillations of axial plant organs.

The tips of roots and shoots commonly show lateral movements as they grow forwards. These occur as both circumnutations (with long periods and large amplitudes) and micronutations (with short periods and small amplitudes). Their properties are reviewed, with emphasis on roots, and possible ways in which they could be regulated are discussed. The mechanisms could include long-range controls (for circumnutations) that depend on transmissible signals using steps common to gravitropism, and short-range controls (for micronutations) that operate within the elongation zone. The former are a property of the apex as a whole, while the latter may be confined to localized groups of cells. Simulation of nutations is presented with a view to isolating key physiological processes. However, this approach is limited by the current inadequate understanding of the growth mechanisms involved.     

New considerations of the oxygen effects in radiation biology.

The original demonstration in the bacterial spore of the multiple actions of oxygen in modifying the responses of cells to ionizing radiation has now been verified in mammalian systems, pointing up the need for separate inquiry into each of the several components if the responses of mammalian cells are to be understood. We have provided physico-chemical explanations for only two of the four (at least) oxygen elements recognized in the spore: the reaction of diatomic oxygen with organic free radicals in dry spores; and an action of the hydroxy radical (OH) in spores at low [O2]. This paper will discuss newly recognized features of the oxygen responses that could reveal the nature of the other components. It will examine modulation of response by oxides of nitrogen, the similarities and differences among them and explanations for them that suggest further experiment, the importance of concentration of additive in determining both quantity and quality of response, and a general model that explains sensitizer action in terms of inhomogeneous chemistry.     

Cell biology of plant gravity sensing.

The debate about whether gravity sensing relies upon statoliths (amyloplasts that sediment) has intensified with recent findings of gravitropism in starchless mutants and of claims of hydrostatic gravity sensing. Starch and significant plastid sedimentation are not necessary for reduced sensing in mutant roots, but plastids might function here if there were a specialized receptor for plastid mass e.g. in the ER. Alternatively, components in addition to amyloplasts might provide mass for sensing. The nucleus is dense and its position is regulated, but no direct data exist for its role in sensing. If the weight of the protoplast functioned in sensing, why would there be specific cytological specializations favoring sedimentation rather than cell mass? Gravity has multiple effects on plants in addition to gravitropism. There may be more than one mechanism of gravity sensing.     

Machine vision monitoring of plant health.

Techniques and algorithms to detect and diagnose disorders in plants grown in a controlled environment have been developed. A video camera senses features of plants which are indicative of disorders. Images are calibrated for size and color variations by using calibration templates. Different image segmentation techniques for separating object from background, have been implemented. Plant size and color properties have been investigated, temporal, spectral and spatial variation of leaves were extracted from the segmented images. Neural network and statistical classifiers were used to determine plant condition.     

Microdosimetric considerations of effects of heavy ions on E. coli K-12 mutants.

The inactivation cross sections of E. coli K-12 recombination-deficient mutants, JC1553 (recA) and AB2470 (recB), for several MeV/u alpha-particles and N ions have been successfully analyzed by Katz's target theory in which radiosensitivity parameter E0 is assumed to be LET independent and equal to D37 for gamma-rays. For E. coli K-12 wild type, AB1157 (rec+, uvr+), however, it is impossible to interpret the inactivation cross section data by an LET-independent E0-value. In the latter case, as in the case of B. subtilis spore, it is necessary to assume that the radiosensitivity of the target for the core of a heavy ion is higher than that for delta-electrons. As well as Waligorski, Hamm and Katz's dose, the dose around the trajectory of an ion based on Tabata and Ito's energy deposition algorithm for electrons has been used in the course of analysis.     

Effects of air current speed on gas exchange in plant leaves and plant canopies.

To obtain basic data on adequate air circulation to enhance plant growth in a closed plant culture system in a controlled ecological life support system (CELSS), an investigation was made of the effects of the air current speed ranging from 0.01 to 1.0 m s-1 on photosynthesis and transpiration in sweetpotato leaves and photosynthesis in tomato seedlings canopies. The gas exchange rates in leaves and canopies were determined by using a chamber method with an infrared gas analyzer. The net photosynthetic rate and the transpiration rate increased significantly as the air current speeds increased from 0.01 to 0.2 m s-1. The transpiration rate increased gradually at air current speeds ranging from 0.2 to 1.0 m s-1 while the net photosynthetic rate was almost constant at air current speeds ranging from 0.5 to 1.0 m s-1. The increase in the net photosynthetic and transpiration rates were strongly dependent on decreased boundary-layer resistances against gas diffusion. The net photosynthetic rate of the plant canopy was doubled by an increased air current speed from 0.1 to 1.0 m s-1 above the plant canopy. The results demonstrate the importance of air movement around plants for enhancing the gas exchange in the leaf, especially in plant canopies in the CELSS.     

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.     

The function of calcium in plant graviperception.

The fundamental question of gravitational biology is how do plants perceive a gravity. Recent experimental results have demonstrated that Ca second-messenger system has an essential role in induction of graviresponsiveness. Our data, that stimuli of various nature cause a rise of hyaloplasm Ca level revealed by means of pyroantimonate method, as well as complete inhibition of the gravitropism in roots of pea seedlings, provide indirect but consistent evidence of this role of Ca ions. A possible explanation for these results is that they may be due to an unbalanced and undirectional influx of Ca ions in statocytes from cell walls or from intracellular Ca stores, while in the presence of the Earths 1 g vector, this process occurs directionally, along this vector. It is possible that a target for the gravity stimulus is the flux mechanism of Ca to statocytes, including participation of the phosphatidylinositol system and calmodulin. The data that have become available from space flight experiments will be reviewed and an attempt will be made to compare these results with ground-based observations.     

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).     

Planetary protection considerations for MarsNet and Mars sample return missions.

The ESA MarsNet mission proposal consists most probably of a trio of Mars landers. These landers each contain a variety of scientific equipment. The network of stations demands for a definition of its planetary protection requirements. With respect to the MarsNet mission only forward contamination problems will be considered. Future involvement of European efforts in planetary exploration including sample returns will also raise the problem of back contamination. A tradeoff study for the overall scientific benefit with respect to the approximative cost is necessary. Planetary protection guide-lines will be proposed by an interdisciplinary and international board of experts working in the fields of both biology and planetary science. These guide-lines will have to be flexible in order to be modified with respect to new research results, e.g. on adaptation of microorganisms to extreme (space) conditions. Experiments on the survival of microorganisms at conditions of simulated Mars surface and subsurface will have to be conducted in order to obtain a baseline data collection as a reference standard for future guide-lines.     

Instrumentation for plant health and growth.

Comprehensive spectroscopic monitoring of plant health and growth in bioregenerative life support system environments is possible using a variety of spectrometric technologies. Absorption spectrometry and atomic emission spectrometry in combination allow for direct, on-line, reagentless monitoring of plant nutrients from nitrate and potassium to micronutrients such as copper and zinc. Fluorometric spectrometry is ideal for the on-line detection, identification and quantification of bacteria and fungi. Liquid Atomic Emission Spectrometry (LAES) is a new form of spectrometry that allows for direct measurement of atomic emission spectra in liquids. An electric arc is generated by a pair of electrodes in the liquid to provide the energy necessary to break molecular bonds and reduce the substance to atomic form. With a fiber probe attached to the electrodes, spectral light can be transmitted to a photodiode array spectrometer for light dispersion and analysis. Ultraviolet (UV) absorption spectrometry is a long-established technology, but applications typically have required specific reagents to produce an analyte-specific absorption. Nitrate and iron nutrients have native UV absorption spectra that have been used to accurately determine nutrient concentrations at the +/- 5% level. Fluorescence detection and characterization of microbes is based upon the native fluorescent signatures of most microbiological species. Spectral and time-resolved fluorometers operating with remote fiber-optic probes will be used for on-line microbial monitoring in plant nutrient streams.     

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.     

Theoretical model of HZE particle fragmentation by hydrogen targets.

The fragmenting of high energy, heavy ions (HZE particles) by hydrogen targets is an important, physical process in several areas of space radiation research. In this work quantum mechanical optical model methods for estimating cross sections for HZE particle fragmentation by hydrogen targets are presented. The cross sections are calculated using a modified abrasion-ablation collision formalism adapted from a nucleus-nucleus collision model. Elemental and isotopic production cross sections are estimated and compared with report measurements for the breakup of neon, sulphur, and iron, nuclei at incident energies between 400 and 910 MeV/nucleon. Good agreement between theory and experiment is obtained.     

Radiosensibility of higher plant seeds after space flight.

The influence of long-term storage of higher plant seeds under space flight conditions (49 to 827 days) on their radiosensibility was studied in the experiments on the orbital stations Salyut 6 and 7. Short-term storage has been proved to have no effect on radiosensitivity of Crepis capillaris seeds. Only in the case of maximal exposure duration the frequency of chromosome aberrations in post-flight irradiated seeds significantly exceeded the chromosome aberration frequency in the ground-based irradiated control. A statistically significant increase in the number of cells with multiple chromosome aberrations was also observed in this experiment. After gamma-irradiation of Arabidopsis thaliana seeds the germinating ability and survival rate of plants decreased depending on the duration of seed storage. Flight-exposed seeds were more sensitive to irradiations with respect to these parameters. A statistically significant increase in the frequency of recessive lethal mutations was observed only in two experiments of long exposure duration.     

Evidence for electrotropism in some plant species.

The ever-present global Atmospheric Electrical Field (AEF) is used by many plant species. There are many natural habitats with electrotropic plants and habitats with no AEF. The plants growing there are not electrotropic, like the plants growing under the canopies of the trees or the Arecibo radio telescope. Examples are given of different plants which belong to one or the other class, and the criteria how to distinguish them. In addition to natural habitat observations, laboratory experiments were run in search of the sensitivity of electrotropic effect to different electric field intensities. During a few years, it was established that in very strong fields (of the order of 1 MV/m) all plants respond immediately to the field. This type of reaction is due to the Coulomb forces, but electrotropism depends on electric field interaction with ions. The "reference field" (130 V/m) was always used with stronger fields in the several kV/m range which enhance plant growth rate and size similar to plant growth hormones. Surprising effects were also observed with reversed and horizontal field polarity. In conclusion electrotropic plants deprived of the electrical field do not develop as expected, as can be seen in Biosphere 2. This is an instructive example of what happens when we forget to provide the plants with this vital natural environmental factor. Electrical fields of different intensity, directions and configurations are cheap and easy to generate.