Growth protocols for etiolated soybeans germinated within BRIC-60 canisters under spaceflight conditions.

As part of the GENEX (Gene Expression) spaceflight experiment, protocols were developed to optimize the inflight germination and subsequent growth of 192 soybean (Glycine max cv McCall) seeds during STS-87. We describe a method which provided uniform growth and development of etiolated seedlings while eliminating root and shoot restrictions for short-term (4-7 day) experiments. Final seedling growth morphologies and the gaseous CO2 and ethylene levels present both on the last day in space and at the time of recovery within the spaceflight and ground control BRIC-60 canisters are presented.     

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.     

双剪连接件及双耳连接耳片疲劳寿命估算的逐次累计求和算法

为了研究复杂连接件疲劳失效机理和估算其裂纹形成和扩展寿命,在双剪连接件和双耳连接耳片疲劳试验的基础上,首先,通过扫描电子显微镜（SEM）分析,研究了其破坏模式和机理,并利用断口定量反推技术判读了疲劳裂纹形成与扩展寿命。然后,根据应力严重系数法,建立了复杂连接件疲劳性能S-N-L（疲劳应力-寿命-应力严重系数）曲面;利用该曲面,发展了复杂连接件疲劳裂纹形成寿命估算算法;基于断裂力学理论,推导出裂纹扩展长度与扩展角度公式,建立了疲劳裂纹扩展寿命估算的累计求和算法。最后,运用寿命估算方法,估算了双剪连接件的疲劳裂纹形成寿命、双剪连接件和双耳连接耳片的裂纹扩展寿命,预测结果与断口判读结果吻合良好。本文寿命估算方法为复杂连接件疲劳定寿提供了理论依据。      

Biological nitrogen fixation under primordial Martian partial pressures of dinitrogen.

Early Earth and early Mars were similar enough such that past geochemical and climatic conditions on Mars may have also been favorable for the origin of life. However, one of the most striking differences between the two planets was the low partial pressure of dinitrogen (pN2) on early Mars (18 mb). On Earth, nitrogen is a key biological element and in many ecosystems the low availability of fixed nitrogen compounds is the main factor limiting growth. Biological fixation of dinitrogen on Earth is a crucial source of fixed nitrogen. Could the low availability of dinitrogen in the primordial Martian atmosphere have prevented the existence, or evolution of Martian microbiota? Azotobacter vinelandii and Azomonas agilis were grown in nitrogen free synthetic medium under various partial pressures of dinitrogen ranging from 780-0 mb (total atmosphere=1 bar). Below 400 mb the biomass, cell number, and growth rate decreased with decreasing pN2. Both microorganisms were capable of growth at a pN2 as low as 5 mb, but no growth was observed at a pN2 < or = 1 mb. The data appear to indicate that biological nitrogen fixation could have occurred on primordial Mars (pN2=18 mb) making it possible for a biotic system to have played a role in the Martian nitrogen cycle. It is possible that nitrogen may have played a key role in the early evolution of life on Mars, and that later a lack of available nitrogen on that planet (currently, pN2=0.2 mb) may have been involved in its subsequent extinction.     

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.     

Super-optimal CO2 reduces wheat yield in growth chamber and greenhouse environments.

Seven growth chamber trials (six replicate trials using 0.035, 0.12, and 0.25% CO2 in air and one trial using 0.12, 0.80, and 2.0% CO2 in air) and three replicate greenhouse trials (0.035, 0.10, 0.18, 0.26, 0.50, and 1.0% CO2 in air) compare the effects of super-optimal CO2 on the seed yield, harvest index, and vegetative growth rate of wheat (Triticum aestivum L. cvs. USU-Apogee and Veery-10). Plants in the growth chamber trials were grown hydroponically under fluorescent lamps, while the greenhouse trials were grown under sunlight and high pressure sodium lamps and in soilless media. Plants in the greenhouse trials responded similarly to those in the growth chamber trials; maximum yields occurred near 0.10 and 0.12% CO2 and decreased significantly thereafter. This research indicates that the toxic effects of elevated CO2 are not specific to only one environment and has important implications for the design of bio-regenerative life support systems in space, and for the future of terrestrial agriculture.     

Efficacy of an ototoxic aminoglycoside (gentamicin) on the differentiation of the inner ear of cichlid fish.

Previous investigations revealed that the growth of fish inner ear otoliths depends on the amplitude and the direction of gravity, thus suggesting the existence of a (negative) feedback mechanism. In the course of these experiments, it was shown that altered gravity both affected otolith size (and thus the provision of the proteinacious matrix) as well as the incorporation of calcium. It is hitherto unknown, as of whether sensory hair cells are involved either in the regulation of otolith growth or in the provision of otolithic material (such as protein or inorganic components) or even both. The ototoxic aminoglycoside gentamicin (GM) damages hair cells in many vertebrates (and is therefore used for the treatment of Meniere's disease in humans). The present study was thus designed to determine as of whether vestibular sensory cells are needed for otolith growth by applying GM in order to induce a (functionally relevant) loss of these cells. Developing cichlid fish Oreochromis mossambicus were therefore immersed in 120 mg/l GM for 10 or 21 days. At the beginning and at the end of the experimental periods, the fish were incubated in the calcium-tracer alizarin complexone (AC). After the experiment, otoliths were dissected and the area grown during GM-exposure (i.e., the area enclosed by the two AC labellings) was determined planimetrically. The results showed that incubating the animals in a GM-solution had no effect on otolith growth, but the development of otolith asymmetry was affected. Ultrastructural examinations of the sensory hair cells revealed that they had obviously not been affected by GM-treatment (no degenerative morphological features observed). Overall, the present results suggest that hair cells are not affected by GM concerning their possible role in (general) otolith growth, but that these cells indeed might have transitionally been impaired by GM resulting in a decreased capacity of regulating otolith symmetry.     

Assessing ecohydrological factors variations and their relationships at different spatio-temporal scales in semiarid area,northwestern China

As a typical semiarid farming-pastoral ecotone sensitive to the environmennt, the Plain of West Liaohe Basin (WLBP) is currently experiencing drastic environmental changes. To identify how environmental change affect vegetation in the WLBP, we analysed spatiotemporal variation characteristics of Ecological environment factors based on monthly and annual air temperature (T), precipitation (P) and Normalized Difference Vegetation Index (NDVI) from 1982 to 2015. And the correlations between them were investigated by correlation analysis (Simple correlation, partial correlation and complex correlation) at temporal and spatial scale. The results showed that: (1) the vegetation growth of the WLBP showed ameliorated trend, with a change rate of 0.004/yr.; (2) P was more sensitive to NDVI than T; (3) and the influence of hydrothermal changes on vegetation growth was more significant than that of the change of single climate factors at time scales; (4) the effects of anthropogenic factors on vegetation change were 75.07% (1982–1993) and 98.08% (1994–2015), respectively. At the temp-special scales, P&T and land use type change (LUCC) were the main climatic and anthropogenic factors that affect vegetation changes, respectively.     

Comparison of aerobically-treated and untreated crop residue as a source of recycled nutrients in a recirculating hydroponic system.

This study compared the growth of potato plants on nutrients recycled from inedible potato biomass. Plants were grown for 105 days in recirculating, thin-film hydroponic systems containing four separate nutrient solution treatments: (1) modified half-strength Hoagland's (control), 2) liquid effluent from a bioreactor containing inedible potato biomass, 3) filtered (0.2 micrometer) effluent, and 4) the water soluble fraction of inedible potato biomass (leachate). Approximately 50% of the total nutrient requirement in treatments 2-4 were provided (recycled) from the potato biomass. Leachate had an inhibitory effect on leaf conductance, photosynthetic rate, and growth (50% reduction in plant height and 60% reduction in tuber yield). Plants grown on bioreactor effluent (filtered or unfiltered) were similar to the control plants. These results indicated that rapidly degraded, water soluble organic material contained in the inedible biomass, i.e., material in leachate, brought about phytotoxicity in the hydroponic culture of potato. Recalcitrant, water soluble organic material accumulated in all nutrient recycling treatments (650% increase after 105 days), but no increase in rhizosphere microbial numbers was observed.     

The nutritional adequacy of a limited vegan diet for a Controlled Ecological Life-Support System.

Purdue University, as well as the Johnson and Kennedy Space Centers and NASA Ames Research Center, are investigating approximately 5-10 plants that will be grown hydroponically to provide not only the energy and nutrients, but also the oxygen for humans habitating in Mars and lunar bases. The growth and nutritional status of rats fed either a control diet (adequate in all macro- and micronutrients) or a strict vegetarian diet consisting of 5 (vegan-5) or 10 (vegan-10) candidate crop species were investigated. In addition, vegan-10 diets were supplemented with mineral and/or vitamin mix at a level similar to the control diets to assess the effect of supplementation on nutrient status. The assessment of inedible plant material as an alternative food source was also investigated. Results of this study demonstrated that consumption of the vegan-10 diet significantly improved weight gain of rats compared to that for rats fed the vegan-5 diet. Mineral supplementation, at a level present in the control diet, to the vegan-10 diet improved growth and nutrient status, but growth was significantly lower compared to the control-fed rats. Inclusion of inedible plant material, high in ash content, improved some indices of nutrient status, without improving growth.     

掺杂纳米硅薄膜的生长特性

采用等离子增强化学气相沉积(PECVD)方法成功的沉积出掺杂(主要是磷、硼)纳米硅薄膜.探讨了各种生长工艺条件对掺杂纳米硅薄膜的结构与性能的影响及其规律.利用高分辨电镜(HREM)、Raman散射等手段对掺入不同杂质后的纳米硅薄膜的微结构进行初步研究,并从实验和理论上对掺杂纳米硅薄膜的生长特性进行了探讨.得出掺杂纳米硅薄膜具有与掺杂非晶硅薄膜和掺杂微晶硅薄膜不同的生长特性,即杂质原子绝大部分是非活性的,只有很少一部分在薄膜中起施主作用.大部分非活性的杂质原子存在于晶粒间界.     

Interaction of growth-determining systems with gravity

The experiments have been carried out with lettuce shoots on board the Salyut-7 orbital station, the Kosmos-1667 biological satellite and under ground conditions at 180° plant inversion. By means of the centrifuge Biogravistat-1M the threshold value of gravitational sensitivity of lettuce shoots has been determined on board the Salyut-7 station. It was found to be equal to 2.9 × 10⁻³g for hypocotyls and 1.5 × 10⁻⁴g for roots. The following results have been received in the experiment performed on board the Kosmos-1667 satellite: a) under microgravity the proliferation of the meristem cells and the growth of roots did not differ from the control; b) the growth of hypocotyls in length was significantly enhanced in microgravity; c) under microgravity transverse growth of hypocotyls (increase in cross sectional area) was significantly increased due to enhancement of cortical parenchyma cell growth. At 180° inversion in Earth's gravity root extension growth and rate of cell division in the root apical meristem were decreased. The determination of DNA-fuchsin value in the nuclei of the cell root apexes showed that inversion affected processess of the cell cycle preceeding cytokinesis.     

Phytochromes play a role in phototropism and gravitropism in Arabidopsis roots.

Phototropism as well as gravitropism plays a role in the oriented growth of roots in flowering plants. In blue or white light, roots exhibit negative phototropism, but red light induces positive phototropism in Arabidopsis roots. Phytochrome A (phyA) and phyB mediate the positive red-light-based photoresponse in roots since single mutants (and the double phyAB mutant) were severely impaired in this response. In blue-light-based negative phototropism, phyA and phyAB (but not phyB) were inhibited in the response relative to the WT. In root gravitropism, phyB and phyAB (but not phyA) were inhibited in the response compared to the WT. The differences observed in tropistic responses were not due to growth limitations since the growth rates among all the mutants tested were not significantly different from that of the WT. Thus, our study shows that the blue-light and red-light systems interact in roots and that phytochrome plays a key role in plant development by integrating multiple environmental stimuli.     

Crop selection for advanced life support systems in the ESA MELiSSA program: Durum wheat (Triticum turgidum var durum)

As part of an ESA MELiSSA investigation into advanced life support (ALS) candidate crop cultivar selection and growth requirements, the University of Guelph’s Controlled Environment Systems Research Facility (CESRF) conducted a case study on growth and development of four durum wheat cultivars (Triticum turgidum var durum) grown hydroponically under controlled conditions in a sealed environment. Cultivars tested were Canadian developed Avonlea, Commander, Eurostar and Strongfield. There were few fundamental differences in durum quality parameters between hydroponically and field grown wheat, however yields of Eurostar and Strongfield exceeded those of field trials by 41% and 87% respectively.     

Germination and elongation of flax in microgravity.

This experiment was conducted as part of a risk mitigation payload aboard the Space Shuttle Atlantis on STS-101. The objectives were to test a newly developed water delivery system, and to determine the optimal combination of water volume and substrate for the imbibition and germination of flax (Linum usitatissimum) seeds in space. Two different combinations of germination paper were tested for their ability to absorb, distribute, and retain water in microgravity. A single layer of thick germination paper was compared with one layer of thin germination paper under a layer of thick paper. Paper strips were cut to fit snugly into seed cassettes, and seeds were glued to them with the micropyle ends pointing outward. Water was delivered in small increments that traveled through the paper via capillary action. Three water delivery volumes were tested, with the largest (480 microliters) outperforming the 400 microliters and 320 microliters volumes for percent germination (90.6%) and root growth (mean=4.1 mm) during the 34-hour spaceflight experiment. The ground control experiment yielded similar results, but with lower rates of germination (84.4%) and shorter root lengths (mean=2.8 mm). It is not clear if the roots emerged more quickly in microgravity and/or grew faster than the ground controls. The single layer of thick germination paper generally exhibited better overall growth than the two layered option. Significant seed position effects were observed in both the flight and ground control experiments. Overall, the design of the water delivery system, seed cassettes and the germination paper strip concept was validated as an effective method for promoting seed germination and root growth under microgravity conditions.     

Changes in spectral reflectance of wheat leaves in response to specific macronutrient deficiency.

In wheat (Triticum aestivum L.) plants, deficiency of an essential element may drastically affect growth, appearance, and most importantly yield. Wheat, the focus of this study, is one of the crops studied in the CELSS program. Information about nutrient deficiencies in crops grown in controlled environment is essential to optimize food productivity. The main objective of this study was to determine whether deficiency of Nitrogen (N), Phosphorus (P), Potassium (K), Calcium (Ca) and Magnesium (M) alters spectral reflectance properties of wheat leaves. Plants were grown in the greenhouse and growth chamber, in a modified Hoagland's nutrient solution. Spectral reflectance of fully expanded wheat leaves from 280 to 1100 nm, nutrient concentrations (N, P, K, and Ca) and chlorophyll (Chl) were determined when deficiency symptoms were first evident (approximately 6-7 weeks). Chlorophyll content and fresh and dry weight, were used to assess the severity of the nutrient stress. All nutrient deficiencies affected chlorophyll content and generally increased reflectance in the visible (VIS) 400-700 nm and infrared (IR) 700-1100 nm ranges. Magnesium and nitrogen deficiencies had the most pronounced effect on chlorophyll concentration height, and reflectance. All macronutrient deficiencies tested reduced chlorophyll concentration, increase reflectance in the visible range and caused a shift in the position of the red edge (the point of maximum slope on the reflectance spectrum of vegetation between red and near-infrared wavelengths) toward shorter or longer wavelengths; depending upon the element. In the greenhouse, N and Mg induced the greatest increase in reflectance of 33% and 25% in the VI range and 86% and 53% in the IR range, respectively. However, in the growth chamber, an increase of 97% and 25% occurred in the VI range, and 20% and 33% in the IR range, respectively. In the IR range in the growth chamber, P, K, and Ca deficiency caused a reduction in reflectance (412-770 nm). This research indicates that mineral deficiencies and reflectance are not specific to one environment and could have important implications for the design of CELSS in space, and perhaps the future of terrestrial agriculture.     

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.     

Effect of hypergravity on carboanhydrase reactivity in inner ear ionocytes of developing cichlid fish.

It has been shown earlier that hypergravity slows down inner ear otolith growth in developing fish. Otolith growth in terms of mineralization mainly depends on the enzyme carboanhydrase (CA), which is responsible for the provision of the pH-value necessary for calcium carbonate deposition. Larval siblings of cichlid fish (Oreochromis mossambicus) were subjected to hypergravity (3 g, hg; 6 h) during development and separated into normally and kinetotically swimming individuals following the transfer to 1 g (i.e., stopping the centrifuge; kinetotically behaving fish performed spinning movements). Subsequently, CA was histochemically demonstrated in inner ear ionocytes (cells involved in the endolymphatic ion exchange) and enzyme reactivity was determined densitometrically. It was found that both the total macular CA-reactivity as well as the difference in reactivities between the left and the right maculae (asymmetry) were significantly lower (1) in experimental animals as compared to the 1 g controls and (2) in normally swimming hg-animals as compared to the kinetotically behaving hg-fish. The results are in complete agreement with earlier studies, according to which hypergravity induces a decrease of otolith growth and the otolithic calcium incorporation (visualized using the calcium-tracer alizarin complexone) of kinetotically swimming hg-fish was higher as compared to normally behaving hyper-g animals. The present study thus strongly supports the concept that a regulatory mechanism, which adjusts otolith size and asymmetry as well as otolithic calcium carbonate incorporation towards the gravity vector, acts via activation/deactivation of macular CA.     

Characterizing parameters of Jatropha curcas cell cultures for microgravity studies

Jatropha (Jatropha curcas) is a tropical perennial species identified as a potential biofuel crop. The oil is of excellent quality and it has been successfully tested as biodiesel and in jet fuel mixes. However, studies on breeding and genetic improvement of jatropha are limited. Space offers a unique environment for experiments aiming at the assessment of mutations and differential gene expression of crops and in vitro cultures of plants are convenient for studies of genetic variation as affected by microgravity. However, before microgravity studies can be successfully performed, pre-flight experiments are necessary to characterize plant material and validate flight hardware environmental conditions. Such preliminary studies set the ground for subsequent spaceflight experiments. The objectives of this study were to compare the in vitro growth of cultures from three explant sources (cotyledon, leaf, and stem sections) of three jatropha accessions (Brazil, India, and Tanzania) outside and inside the petriGAP, a modified group activation pack (GAP) flight hardware to fit petri dishes. In vitro jatropha cell cultures were established in petri dishes containing a modified MS medium and maintained in a plant growth chamber at 25 ± 2 °C in the dark. Parameters evaluated were surface area of the explant tissue (A), fresh weight (FW), and dry weight (DW) for a period of 12 weeks. Growth was observed for cultures from all accessions at week 12, including subsequent plantlet regeneration. For all accessions differences in A, FW and DW were observed for inside vs. outside the PetriGAPs. Growth parameters were affected by accession (genotype), explant type, and environment. The type of explant influenced the type of cell growth and subsequent plantlet regeneration capacity. However, overall cell growth showed no abnormalities. The present study demonstrated that jatropha in vitro cell cultures are suitable for growth inside PetriGAPs for a period of 12 weeks. The parameters evaluated in this study provide the basic ground work and pre-flight assessment needed to justify a model for microgravity studies with jatropha in vitro cell cultures. Future studies should focus on results of experiments performed with jatropha in vitro cultures in microgravity.     

Strong magnetic field effect on the dissolution process of tetragonal lysozyme crystals.

Either a homogeneous or inhomogeneous magnetic field has been known to dampen the protein crystal growth. To date the mechanism is not clear. However, it was generally proposed that the magnetic field may dampen the convection in the solution, resulting in a reduced crystal growth rate and possibly a good crystal quality, similar to the case of protein crystal growth in space. To understand the mechanism of the magnetic field effect on protein crystal growth, further explorations on the magnetic field effect on protein solution, on the processes of crystal growth and dissolution, and on different crystallization (solution) systems, should be valuable. In this paper we present our recent efforts to study magnetic field effects on the dissolution processes of tetragonal lysozyme crystals under a strong magnetic field. A layer of oriented tetragonal lysozyme crystals was prepared under a temperature gradient and magnetic field, after that the crystals were dissolved by increasing the temperature of the solution. The lysozyme molecules will diffuse upwards due to the steep concentration gradient at the lower side of the cell caused by the dissolution. The evolution of the concentration in the solution was measured in-situ using a Mach-Zehnder interferometer. The results confirmed that the dissolution process of the crystals was slowed by the magnetic field. Judging from the concentration evolution versus time at different positions in the solution, we concluded that the apparent diffusion coefficient of lysozyme molecules was decreased by the magnetic field. The results were discussed using a suspended crystal model in the initial dissolution stage.