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At least 91 records · Page 5

Polar distribution of annexin-like proteins during phytochrome-mediated initiation and growth of rhizoids in the ferns Dryopteris and Anemia

Although the calcium requirement of phytochrome-mediated fern spore germination and early rhizoid growth is well established, the calcium-binding proteins that serve as transducers for these responses are not known. Here we report the presence of annexin-like proteins in germinating spores of Dryopteris filix-mas (L.) Schott and Anemia phyllitidis (L.) Sw. and evidence that they may be important participants in early photomorphogenic changes in gametophytes. Immunolocalization and immunoblot assays of these proteins were carried out using polyclonal antibodies raised either against a 35-kDa annexin-like protein from pea or against anchorin CII from chicken. Western-blot analysis showed that crude protein extracts obtained from both species after red-light treatment contained two cross-reactive protein bands with molecular weights around 70 kDa. These proteins were annexin-like in that they bound to a phosphatidylserine affinity column in a calcium-dependent fashion. Using this column, two protein bands around 70 kDa, i.e. 67 and 73 kDa, were partially purified together with proteins at 36 kDa and a doublet at 54 kDa. Proteins of these latter molecular weights are suggested to be members of the annexin family, but no cross-reactivity could be found between these and the two antibodies used in our investigations. Immunodetectable levels of these proteins were observed only after light-mediated induction of spore germination. Imaging of the immuno-localization patterns observed with both antibodies showed that the annexin-like proteins are concentrated at the extreme tips of the rhizoids in D. filix-mas and A. phyllitidis during rhizoid initiation and all stages of elongation. We suggest that these proteins may play a major role in the tip-oriented exocytosis events that are critical for the initiation and growth of fern rhizoids.

NASA Discipline Number 40-50↗

Influence of Hydroponically Grown Hoyt Soybeans and Radiation Encountered on Mars Missions on the Yield and Quality of Soymilk and Tofu

Soybeans were chosen for hmar and planetary missions due to their nutritive value and ability to produce oil and protein for further food applications. However, soybeans must be processed into foods prior to crew consumption. Wilson et al. (2003) raised questions about (1) the influence of radiation (on germination and functional properties) that the soybeans would be exposed to during bulk storage for a Mars mission, and (2) the impact of using hydroponically grown versus field grown soybeans on the yield and quality of soyfoods. The influence of radiation can be broken down into two components: (A) affect of surface pasteurization to ensure the astronauts safety from food-borne illnesses (a Hazard Analysis Critical Control Point), and (B) affect of the amount of radiation the soybeans receive during a Mars mission. Decreases in the amount of natural antioxidants and free radical formation and oxidation induced changes in the soybean (lipid, protein, etc.) will influence the nutritional value, texture, quality, and safety of soyfoods made from them. The objectives of this project are to (1) evaluate the influence of gamma and electron beam radiation on bulk soybeans (HACCP, CCP) on the microbial load, germination, ease of processing, and quality of soymilk and tofu; (2) provide scale up and mass balance data for Advanced Life Support subsystems including Biomass, Solid Waste Processing, and Water Recovery Systems; and (3) to compare Hoyt field grown to hydroponically grown Hoyt soybeans for soymilk and tofu production. The soybean cultivar Hoyt, a small standing, high protein cultivar that could grow hydroponically in the AIMS facility on Mars) was evaluated for the production of soymilk and tofu. The quality and yield of the soymilk and tofu from hydroponic Hoyt, was compared to Vinton 81 (a soyfood industry standard), field Hoyt, IA 2032LS (lipoxygenase-free), and Proto (high protein and antioxidant potential). Soymilk and tofu were produced using the Japanese method. The soymilk was coagulated with calcium sulfate dihydrate. Soybeans and tofu were evaluated using chemical, microbial, and instrumental sensory methods. The surface radiation of whole dry soybeans using electron beam or gamma rays at 10 or 30 kGy did provide microbial safety for the astronauts. However, these doses caused oxidative changes that resulted in tofu with rancid aroma, darkening of the tofu, lower tofu yields, more solid waste, and loss of the ability of the seeds to germinate. While lower doses may reduce these problems, we lose the ability to insure microbial safety (cross-contamination) of bulk soybeans for the astronauts. Counter measures could include vacuum packaging, radiating under freezing conditions. A No Effect Dose for food quality, below 10 kGy needs to be determined. Better estimates of the radiation that the food will be exposed to need to determined and shared. Appropriate shielding for the food as well as the astronauts needs to be developed. The Hoyt soybean did not provide a high yielding, high quality tofu. A new small scale system for evaluating soybeans was developed using 50 g quantities of soybeans.

Wilson, Lester A.↗

Irradiating of Bulk Soybeans: Influence on Their Functional and Sensory Properties for Soyfood Processing

Soybeans were chosen for lunar and planetary missions, where soybeans will be supplied in bulk or grown locally, due to their nutritive value and ability to produce oil and protein for further food applications. However, soybeans must be processed into foods prior to consumption. Radiation that soybeans would be exposed to during bulk storage prior to and during a Mars mission may influence their germination and functional properties. The influence of radiation includes the affect of surface pasteurization to ensure the astronauts safety from food-borne illnesses (HACCP, CCP), and the affect of the amount of radiation the soybeans receive during a Mars mission. Decreases in the amount of natural antioxidants free radical formation, and oxidation-induced changes in the soybean will influence the nutritional value, texture, color, and aroma of soyfoods. The objective of this study was to determine the influence of pasteurization and sterilization surface radiation on whole soybeans using gamma and electron beam radiation. The influence of 0, 1, 5, 10, and 30kGy on microbial load, germination rate, ease of processing, and quality of soymilk and tofu were determined. Surface radiation of whole dry soybeans using electron beam or gamma rays from 1-30kGy did provide microbial safety for the astronauts. However, the lower dose levels had surviving yeasts and molds. These doses caused oxidative changes that resulted in soymilk and tofu with rancid aromas. GC-MS of the aroma compounds using SPME Headspace confirmed the presence of lipid oxidation compounds. Soybean germination ability was reduced as radiation dosage increased. While lower doses may reduce these problems, the ability to insure microbial safety of bulk soybeans will be lost. Counter measures could include vacuum packaging, nitrogen flushing, added antioxidants, and radiating under freezing conditions. Doses below 1kGy need to be investigated further to determine the influence of the radiation encountered during Mars missions.

Chia, Chiew-Ling↗

Adenosine Monophosphate-Based Detection of Bacterial Spores

A method of rapid detection of bacterial spores is based on the discovery that a heat shock consisting of exposure to a temperature of 100 C for 10 minutes causes the complete release of adenosine monophosphate (AMP) from the spores. This method could be an alternative to the method described in the immediately preceding article. Unlike that method and related prior methods, the present method does not involve germination and cultivation; this feature is an important advantage because in cases in which the spores are those of pathogens, delays involved in germination and cultivation could increase risks of infection. Also, in comparison with other prior methods that do not involve germination, the present method affords greater sensitivity. At present, the method is embodied in a laboratory procedure, though it would be desirable to implement the method by means of a miniaturized apparatus in order to make it convenient and economical enough to encourage widespread use.

Kern, Roger G.↗

Plasma Sanitization of Radish Seeds for International Space Station Growth Experiments

Plant growth in space has long been a topic of key interest to enable sustained human presence. Seeds are common sources for microbial growth and must be sanitized before being sent to space. Current disinfection methods use toxic chemicals that require careful handling and dosing or are ineffective against certain seed types. Low temperature plasma provides an alternative to traditional methods. Plasma produces reactive species and ultraviolet light that reduce contaminants on seeds. NASA’s Kennedy Space Center has explored the treatment of radish seeds with plasma for sanitization and germination. Radio Frequency (RF) and Alternating Current (AC) plasma sources provided continuous treatment of radish seeds in a low-pressure and atmospheric environment, respectively. The seeds were analyzed to compare the presence of bacteria, fungi, and the germination rate. Comparisons between direct and indirect treatments, power supplies, treatment times, and treatment pressures showed the importance of proper dosing of reactive species. Higher powers and longer treatment times have shown larger reductions in microbial contaminants. However, extended direct exposure to high powered plasma can sometimes lead to a reduction in germination rates. Overall, plasma has shown promise as a viable alternative to chemical disinfection techniques. *Work supported by NASA's Kennedy Space Center Engineering and Safety Operations Independent Research and Technology Development.

K W Engeling↗

The Effects of Plasma Application on Radish Seeds with Implications for Space Crop Production

In extended space missions, the astronaut diet will consist mostly of prepackaged foods. This could result in nutritional deficiencies due to the gradual breakdown of certain vitamins. To address this deficiency, fresh produce must be grown from seed during spaceflight. Stored seeds, however, can be vulnerable to microbial contamination which could jeopardize plant health and crop food safety. To alleviate this concern, the current practice is to sanitize seeds on the ground before spaceflight to the International Space Station (ISS). Methods of seed sanitization include alcohol soaking and chlorine gas fumigation, which have harmful effects on the environment and human health. Plasma application is a new sanitization approach that avoids these negative side effects while potentially elevating germination rate and improving growth rate; yet plasma application requires specific exposure time, power, and pressure to achieve these benefits. In the present study, Raphanus sativus ‘Cherry Belle’ radish seeds were exposed to either low pressure (Diener) plasma or atmospheric pressure plasma for varying increments of time (30s-1200s). Data collected includes immediate germination rate (viability) and microbial log reduction. Additionally, seeds were stored for later germination rate testing. Microbial assay controls have highlighted variability in initial microbial load between individual seeds, and further work is being performed to determine the standard microbial load of an untreated seed. In some samples, a negative log reduction was observed after plasma treatment encouraging further study to determine if the plasma is perforating the seed coat and releasing endophytic microbes. Treatment with Diener plasma shows promising microbial log reduction, but lower viability. In contrast, treatment with atmospheric pressure plasma offers high viability, but poor microbial log reduction. Further study will aim at determining whether plasma treatment is effective at not only sanitization, but sterilization. This research was funded by a NASA grant at the Kennedy Space Center.

Plasma↗

The Effects of Plasma Application on Radish Seeds with Implications for Space Crop Production

In extended space missions, the astronaut diet will consist mostly of prepackaged foods. This could result in nutritional deficiencies due to the gradual breakdown of certain vitamins. To address this deficiency, fresh produce must be grown from seed during spaceflight. Stored seeds, however, can be vulnerable to microbial contamination which could jeopardize plant health and crop food safety. To alleviate this concern, the current practice is to sanitize seeds on the ground before spaceflight to the International Space Station (ISS). Methods of seed sanitization include alcohol soaking and chlorine gas fumigation, which have harmful effects on the environment and human health. Plasma application is a new sanitization approach that avoids these negative side effects while potentially elevating germination rate and improving growth rate; yet plasma application requires specific exposure time, power, and pressure to achieve these benefits. In the present study, Raphanus sativus ‘Cherry Belle’ radish seeds were exposed to either low pressure (Diener) plasma or atmospheric pressure plasma for varying increments of time (30s-1200s). Data collected includes immediate germination rate (viability) and microbial log reduction. Additionally, seeds were stored for later germination rate testing. Microbial assay controls have highlighted variability in initial microbial load between individual seeds, and further work is being performed to determine the standard microbial load of an untreated seed. In some samples, a negative log reduction was observed after plasma treatment encouraging further study to determine if the plasma is perforating the seed coat and releasing endophytic microbes. Treatment with Diener plasma shows promising microbial log reduction, but lower viability. In contrast, treatment with atmospheric pressure plasma offers high viability, but poor microbial log reduction. Further study will aim at determining whether plasma treatment is effective at not only sanitization, but sterilization. This research was funded by a NASA grant at the Kennedy Space Center.

Space Crop Production↗

Influence of gravity and light on the developmental polarity of Ceratopteris richardii fern spores

The polarity of germinating single-celled spores of the fern Ceratopteris richardii Brogn. is influenced by gravity during a time period prior to the first cellular division designated a "polarity-determination window". After this window closes, control of polarity is seen in the downward (with respect to gravity) migration of the nucleus along the proximal face of the spore and the subsequent downward growth of the primary rhizoid. When spores are germinated on a clinostat the direction of nuclear migration and subsequent primary rhizoid growth is random. However, in each case the direction of nuclear migration predicts the direction of rhizoid elongation. Although it is the most obvious movement, the downward migration is not the first movement of the nucleus. During the polarity-determination window, the nucleus moves randomly within a region centered behind the trilete marking. While the polarity of many fern spores has been reported to be controlled by light, spores of C. richardii are the first documented to have their polarity influenced by gravity. Directional white light also affects the polarity of these spores, but this influence is slight and is secondary to that of gravity.

Non-NASA Center↗

Camelina circRNA landscape: Implications for gene regulation and fatty acid metabolism

Abstract Circular RNAs (circRNAs) are closed‐loop RNAs forming a covalent bond between their 3′ and 5′ ends, the back splice junction (BSJ), rendering them resistant to exonucleases and thus more stable compared to linear RNAs. Identification of circRNAs and distinction from their cognate linear RNA is only possible by sequencing the BSJ that is unique to the circRNA. CircRNAs are involved in the regulation of their cognate RNAs by increasing transcription rates, RNA stability, and alternative splicing. We have identified circRNAs from C. sativa that are associated with the regulation of germination, light response, and lipid metabolism. We sequenced light‐grown and etiolated seedlings after 5 or 7 days post‐germination and identified a total of 3447 circRNAs from 2763 genes. Most circRNAs originate from a single homeolog of the three subgenomes from allohexaploid camelina and correlate with higher ratios of alternative splicing of their cognate genes. A network analysis shows the interactions of select miRNA:circRNA:mRNAs for regulation of transcript stabilities where circRNA can act as a competing endogenous RNA. Several key lipid metabolism genes can generate circRNA, and we confirmed the presence of KASII circRNA as a true circRNA. CircRNA in camelina can be a novel target for breeding and engineering efforts.

Utley, Delecia [Department of Plant and Microbial ↗

Data for Expression of a Bacterial Trehalose 6-Phosphate Synthase Gene otsA in Camelina sativa Seeds Promotes the Channelling of Carbon Towards Oil Accumulation

Improving seed oil yield is essential for developing Camelina sativa as a sustainable biofuel crop. Fatty acid synthesis depends on the production of acetyl-CoA from photosynthetically derived sugars. Trehalose 6-phosphate (T6P), a proxy for sucrose availability, can link sugar status to plant growth and development. Synthesised by trehalose 6-phosphate synthase (TPS) from UDP-glucose and glucose-6-phosphate, T6P plays a regulatory role in metabolism. Our previous studies on Arabidopsis transgenic lines constitutively expressing the E. coli otsA (encoding TPS) showed increased T6P levels and seed triacylglycerol, along with stunted growth. In the present study we express otsA in camelina under the control of a seed-specific Phaseolin promoter. Seeds of the resulting transgenic lines accumulated high levels of T6P, and a 15%–20% increase in total fatty acids and triacylglycerol compared to wild-type. Molecular analysis showed the transgenic seeds had reduced SnRK1 activity, elevated WRI1 protein levels, and increased the levels of WRI1 and its target genes, along with enhanced rates of fatty acid synthesis that increased seed weights relative to wild type. Notably, the increase in oil did not affect seed protein levels but did reduce the soluble metabolite fraction. Crucially, seed-specific expression of otsA mitigated the growth defects associated with constitutive otsA expression, and the transgenic lines showed normal seed development and germination. These findings demonstrate that targeted T6P modulation via seed-specific otsA expression is an effective metabolic engineering strategy to boost oil production in camelina and potentially in other oilseed crops and bioenergy crops such as energycane, sorghum and miscanthus.

Lipids↗

Degradable Biocomposite Thermoplastic Polyurethanes

In this project, the team developed tough and degradable biocomposite thermoplastic polyurethanes (TPUs) by incorporating bacterial spores into TPUs as a biofunctional living filler. The team screened various bacteria and selected the Bacillus subtilis ATCC 6633 strain as the final candidate, primarily due to its genomic availability, sporulation ability and TPU assimilation activity. The heat-shock tolerance of ATCC 6633 spores was further improved through evolutionary engineering via Adaptive Laboratory Evolution (ALE), demonstrating a 17.7-fold enhanced germination efficiency post heat-shock treatment compared to the wild-type strain (WT). The team fabricated biocomposite TPUs by incorporating lyophilized powder of heat-shock tolerized (HST) spores during the hot melt extrusion (HME) of TPU at 135 °C. The baseline TPU used in this project is a commercially available soft-grade TPU (BCF45) manufactured by BASF. Colony forming unit (CFU) assays quantified that WT and HST spores in the TPU matrix retained approximately 20% and 100% survivability, respectively, after HME. Tensile testing demonstrated that the spores behaved as a polymer-reinforcing filler, positively affecting the overall tensile properties of the biocomposite TPU. For example, biocomposite TPU with WT and HST spores (BC TPU WT and BC TPU HST , respectively) exhibited up to 25% and 37% improved toughness, respectively, compared to TPU without spores. BC TPU HST showed remarkably improved disintegration in autoclaved compost (92% mass loss in 5 months), which simulated a microbially poor environment for TPU degradation. When compared to TPU without spores (44% mass loss in 5 months) the acceleration of degradation is marked. Respirometry confirmed that 72% of BC TPU HST was biomineralized into CO2 within 6 months, indicating that spores in the biocomposite TPU were germinated by utilizing nutrients in the autoclaved compost, facilitating TPU degradation at the end of the material's life. The team demonstrated the scale-up of biocomposite TPU fabrication using continuous extrusion and injection molding techniques. Processing conditions optimized in a lab-scale microcompounder were successfully transferred to a continuous extruder with a 30-fold increased throughput. Biocomposite TPUs prepared using these industry-relevant processes showed comparable toughness improvements to samples prepared in the lab-scale extruder. Excitingly, following compounding in the pilot-extruder the composite material could be injection molded, while retaining high spore viability and similar toughness improvements. The team also found that spores in biocomposite TPU served as antioxidants, preventing toughness decay during the recycled extrusion of BC TPU HST . Long-term storage tests over one year showed that the addition of spores had no negative effect on the longevity of the TPU. Furthermore, the team demonstrated the fabrication of spore-bearing biocomposite polymers with other polyesters such as PBAT, PLA, and PCL. We obtained promising preliminary data that showed overall toughness improvements for all polymers with spore addition. Finally, life cycle assessment (LCA) and techno-economic analysis (TEA) were carried out, which indicated minimal additional cost of fabrication. Overall, a tough and degradable biocomposite thermoplastic was successfully developed through this project, with all tasks completed successfully, achieving >100% of the objectives.

36 MATERIALS SCIENCE↗

Ecology and thermal inactivation of microbes in and on interplanetary space vehicle components

The mechanism for thermal inactivation of bacterial spores under moist or dry heat was studied. Experimental conditions were established relating to spore loss of heat resistance and loss of optical density as a measure of the rate and extent of germination in spore suspensions. Events occurring during germination were correlated with phase darkening (refractility and non-refractility of spores), stainability characteristics of heat and non-heat treated spores, morphological characteristics, and studies on swelling of spores by an increase in packed cell volume.

Campbell, J. E.↗

Growth and development of plants flown on the STS-3 Space Shuttle mission

Pre-germinated pine seedlings and germinating oat and mung bean seeds were flown on the STS-3 Space Shuttle mission. Overall, the seedlings grew and developed well in space. Some oat and mung bean roots, however, grew upward. Lignin content was slightly lower in flight tissues and protein content was higher.

Cowles, J. R.↗

Growth of plant tissue cultures in simulated lunar soil: Implications for a lunar base Controlled Ecological Life Support System (CELSS)

Experiments to determine whether plant tissue cultures can be grown in the presence of simulated lunar soil (SLS) and the effect of simulated lunar soil on the growth and morphogenesis of such cultures, as well as the effect upon the germination of seeds and the development of seedlings were carried out . Preliminary results on seed germination and seedling growth of rice and calli growth of winged bean and soybean indicate that there is no toxicity or inhibition caused by SLS. SLS can be used as a support medium with supplements of certain major and micro elements.

Venketeswaran, S.↗

Growth of plant tissue cultures in simulated lunar soil: Implications for a lunar base CELSS (Controlled Ecological Life Support System)

Experiments were carried out on plant tissue cultures, seed germination, seedling development and plants grown on Simulated Lunar Soil to evaluate the potential of future development of lunar based agriculture. The studies done to determine the effect of the placement of SLS on tissue cultures showed no adverse effect of SLS on tissue cultures. Although statistically insignificant, SLS in suspension showed a comparatively higher growth rate. Observations indicate the SLS, itself cannot support calli growth but was able to show a positive effect on growth rate of calli when supplemented with MS salts. This positive effect related to nutritive value of the SLS was found to have improved at high pH levels, than at the recommended low pH levels for standard media. Results from seed germination indicated that there is neither inhibitory, toxicity nor stimulatory effect of SLS, even though SLS contains high amounts of aluminum compounds compared to earth soil. Analysis of seeding development and growth data showed significant reduction in growth rate indicating that, SLS was a poor growth medium for plant life. This was confirmed by the studies done with embryos and direct plant growth on SLS. Further observations attributed this poor quality of SLS is due to it's lack of essential mineral elements needed for plant growth. By changing the pH of the soil, to more basic conditions, the quality of SLS for plant growth could be improved up to a significant level. Also it was found that the quality of SLS could be improved by almost twice, by external supply of major mineral elements, directly to SLS.

Venketeswaran, S.↗

Productivity and food value of Amaranthus cruentus under non-lethal salt stress

Experiments were carried out to analyze the effects of increasing salinity stress on growth, photosynthesis, and carbon allocation in the crop plant Amaranthus. Plants were germinated and grown in Hoagland's solution with NaCl concentrations of 0 to 1.0 percent. The limits of total salinity in the plant growth medium are investigated. For Amaranthus cruentus, germination, vegetative growth, flowering, seed development and yield were normal at salinities from 0 to 0.2 percent. Inhibition of these phases increased from o.2 to 0.4 percent salinity and was total above 0.5 percent with 1 percent salinity was lethal to all developmental phases. Onset of growth phases were not affected by salinity. Plants could not be adapted by gradually increasing salinity over days or weeks. Water uptake increased, while photosynthetic CO2 uptake decreased with increasing salinity on a dry weight basis during vegetative growth. Protein levels were unchanged with increasing salinity. Leaf starch levels were lower at salinities of 0.5 percent and above, while stem starch levels were not affected by these salinities. The evidence supports salt inhibition arising frm changes in primary biochemical processes rather than from effects on water relations. While not addressing the toxic effects of specific ions, it suggests that moderate salinity per se need not be a problem in space systems.

Macler, Bruce A.↗

Seeds in space experiment results

Two million seeds of 120 different varieties representing 106 species, 97 genera, and 55 plant families were flown aboard the Long Duration Exposure Facility (LDEF). The seeds were housed on the space exposed experiment developed for students (SEEDS) tray in sealed canister number six and in two small vented canisters. The tray was in the F-2 position. The seeds were germinated and the germination rates and development of the resulting plants compared to the control seed that stayed in Park Seed's seed storage facility. The initial results are presented. There was a better survival rate in the sealed canister in space than in the storage facility at Park Seed. At least some of the seeds in each of the vented canisters survived the exposure to vacuum for almost six years. The number of observed apparent mutations was very low.

Alston, Jim A.↗

Growth, differentiation and development of Arabidopsis thaliana under microgravity conditions (7-IML-1)

The aim of this set of experiments is to quantify the structural and behavioral changes taking place in germinating seeds of the small plant Arabidopsis thaliana. The protocol to be used will involve germination of the seeds in orbit. Their growth will be followed by fixing and photographing samples of microgravity grown and 1 g control seedlings at intervals over 4 days. The different studies which will then be performed can be classified in relation to the parts of the plants involved. The first study will be an examination of the ultrastructure of the root statocytes, the cells containing gravity sensors, to determine whether their development proceeds normally under microgravity conditions. A second study will examine the differences in root and shoot development and orientation between normal wild type seedlings and those of an agravitropic mutant (aux-1) - one that does not respond normally to gravity. A third set of observations will be made on the structural changes occurring during reserve breakdown and utilization in the cells of the cotyledons, the storage organs of the seed. The fourth part of the work will be an examination of the statocytes present in the shoot and their development in microgravity. The final part of the study will give an answer to the debated question of whether the formation of the hypocotyl hook in seedlings is gravity dependent.

Maher, E. P.↗