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Impact of Space Radiation on Plant Seeds: from Arabidopsis thaliana to Crops

One of the major concerns for long-term exploration missions beyond the Earth’s magnetosphere is radiation risk, primarily from solar particle events (SPE) and galactic cosmic rays (GCR). With the goal of manned Mars exploration, the production of fresh food during long-duration space missions provides critical nutritional supplementation and may also benefit astronauts’ behavioral health. However, the effects of space radiation on plants and plant propagules have not been sufficiently investigated and characterized. This is the final report of our project to summarize the findings of the effect of space radiation exposure on plant seeds from morphometrics, nutritional values, to molecular machenisms. In this study, we evaluated the effect of simulated GCR (using dry seeds) or SPE (using hydrated seeds) on seeds of Arabidopsis, Mizuna mustard, ‘Outredgeous’ red romaine lettuce, and ‘Red Robin’ dwarf tomato. Seeds were exposed to various doses of simulated space radiation scenarios, either acutely or at a low dose rate (LDR), using the NASA Space Radiation Laboratory (NSRL) facility at Brookhaven National Lab (BNL). Exposure to simulated GCR or SPE at the levels tested had no significant impact on the germination rate in Arabidopsis and crop seeds; however, GCR reduced the viability of lettuce and tomato seeds. Overall, the morphological changes of the seedlings cultured from irradiated seeds were dose- and ion quality- dependent, with heavier ions causing more severe damage. These changes ranged from cotyledon deformation, shortened root length, smaller seedling size, and other signs of stress, depending on the seed types. Both 40 and 80 cGy (LDR) exposures of GCR or SPE significantly affected tomato early seedling development, delayed tomato fruiting, and reduced the total yield of tomato. Altered nutritional values were also found in edible biomass, especially for the GCR 80 cGy (LDR) groups. Underlying mechanisms were furtherly evaluated using transcriptomic analysis. For both GCR and SPE, 40 cGy showed some effects, but to a much lesser extent compared with 80 cGy, which can be considered as the “maximum permissible exposure” for the seed types we evaluated in this study. The impact of space radiation on seeds potentially affects the ability of plants to adapt to other environmental stresses (e.g. microgravity, water stress, and hardware constraints) as well as susceptibility to plant diseases, which need to be furtherly investigated. This research is funded by NASA’s Human Research Program.

J T Richards↗

Impact of Space Radiation on Plant Seeds: from Arabidopsis thaliana to Crops

One of the major concerns for long-term exploration missions beyond the Earth’s magnetosphere is radiation risk, primarily from solar particle events (SPE) and galactic cosmic rays (GCR). With the goal of manned Mars exploration, the production of fresh food during long-duration space missions provides critical nutritional supplementation and may also benefit astronauts’ behavioral health. However, the effects of space radiation on plants and plant propagules have not been sufficiently investigated and characterized. This is the final report of our project to summarize the findings of the effect of space radiation exposure on plant seeds from morphometrics, nutritiaonal values, to molecular machenisms. In this study, we evaluated the effect of simulated GCR (using dry seeds) or SPE (using hydrated seeds) on seeds of Arabidopsis, Mizuna mustard, ‘Outredgeous’ red romaine lettuce, and ‘Red Robin’ dwarf tomato. Seeds were exposed to various doses of simulated space radiation scenarios, either acutely or at a low dose rate (LDR), using the NASA Space Radiation Laboratory (NSRL) facility at Brookhaven National Lab (BNL). Exposure to simulated GCR or SPE at the levels tested had no significant impact on the germination rate in Arabidopsis and crop seeds; however, GCR reduced the viability of lettuce and tomato seeds. Overall, the morphological changes of the seedlings cultured from irradiated seeds were dose- and ion quality- dependent, with heavier ions causing more severe damage. These changes ranged from cotyledon deformation, shortened root length, smaller seedling size, and other signs of stress, depending on the seed types. Both 40 and 80 cGy (LDR) exposures of GCR or SPE significantly affected tomato early seedling development, delayed tomato fruiting, and reduced the total yield of tomato. Altered nutritional values were also found in edible biomass, especially for the GCR 80 cGy (LDR) groups. Underlying mechanisms were furtherly evaluated using transcriptomic analysis. For both GCR and SPE, 40 cGy showed some effects, but to a much lesser extent compared with 80 cGy, which can be considered as the “maximum permissible exposure” for the seed types we evaluated in this study. The impact of space radiation on seeds potentially affects the ability of plants to adapt to other environmental stresses (e.g. microgravity, water stress, and hardware constraints) as well as susceptibility to plant diseases, which need to be furtherly investigated. This research is funded by NASA’s Human Research Program.

J.T. Richards↗

Nutrition of Antarctic-grown Crops to Supplement the Crew Diet, with Applications for Spaceflight

The availability of fresh produce during longer-duration spaceflight missions is being explored as a countermeasure for human biobehavioral health and performance, including how fresh fruits and vegetables can supplement the crew diet with nutrients that are predicted to become deficient throughout the missions. Crop production in the Veggie and Advanced Plant Habitat vegetable production chambers on the International Space Station (ISS) have tested pick-and-eat crops in spaceflight. However, the limited plant cultivation volume of these chambers has restricted the sample sizes and biomass allocated for nutrition assessment. To gain a better understanding of the nutrient composition of pick-and-eat crops, leafy greens and fruiting plants were grown in the EDEN ISS plant cultivation facility near the German Neumayer Station III in Antarctica from March 2021 to January 2022. Target crops for nutrition sampling aligned with cultivars grown in spaceflight, including ‘Outredgeous’ red romaine lettuce, Mizuna mustard, ‘Red Robin’ dwarf cherry tomato, and NuMex ‘Española Improved’ chili pepper. Plants subsamples were taken at harvest; in the event of multiple harvests from the same plants, subsamples were taken at both the first and final harvests. ‘Outredgeous’ and Mizuna were also grown multiple times throughout the year, allowing multiple grow-outs to be analyzed for potential effects across the growing season. Subsamples were first weighed for fresh mass, dried in a dedicated oven at 70°C for 96 h, weighed again, and stored in air-tight containers inside Neumayer Station III. At the beginning of 2022, the samples were shipped to NASA’s Kennedy Space Center, where they are currently being analyzed with ion chromatography for nutrients of interest in the astronaut diet, including calcium, iron, magnesium, and potassium. This study aims to increase our understanding of how the crops that have successfully grown on the ISS can supplement the crew diet, as well as how other factors like plant age and the number of days the facility has been in operation may impact nutrient concentrations. Such findings can improve the crop selection process and how pick-and-eat crops are cultivated during spaceflight missions.

Jess M. Bunchek↗

Nutrition of Antarctic-grown Crops to Supplement the Crew Diet, with Applications for Spaceflight

The availability of fresh produce during longer-duration spaceflight missions is being explored as a countermeasure for human biobehavioral health and performance, including how fresh fruits and vegetables can supplement the crew diet with nutrients that are predicted to become deficient throughout the missions. Crop production in the Veggie and Advanced Plant Habitat vegetable production chambers on the International Space Station (ISS) have tested pick-and-eat crops in spaceflight. However, the limited plant cultivation volume of these chambers has restricted the sample sizes and biomass allocated for nutrition assessment. To gain a better understanding of the nutrient composition of pick-and-eat crops, leafy greens and fruiting plants were grown in the EDEN ISS plant cultivation facility near the German Neumayer Station III in Antarctica from March 2021 to January 2022. Target crops for nutrition sampling aligned with cultivars grown in spaceflight, including ‘Outredgeous’ red romaine lettuce, Mizuna mustard, ‘Red Robin’ dwarf cherry tomato, and NuMex ‘Española Improved’ chili pepper. Plants subsamples were taken at harvest; in the event of multiple harvests from the same plants, subsamples were taken at both the first and final harvests. ‘Outredgeous’ and Mizuna were also grown multiple times throughout the year, allowing multiple grow-outs to be analyzed for potential effects across the growing season. Subsamples were first weighed for fresh mass, dried in a dedicated oven at 70°C for 96 h, weighed again, and stored in air-tight containers inside Neumayer Station III. At the beginning of 2022, the samples were shipped to NASA’s Kennedy Space Center, where they are currently being analyzed with ion chromatography for nutrients of interest in the astronaut diet, including calcium, iron, magnesium, and potassium. This study aims to increase our understanding of how the crops that have successfully grown on the ISS can supplement the crew diet, as well as how other factors like plant age and the number of days the facility has been in operation may impact nutrient concentrations. Such findings can improve the crop selection process and how pick-and-eat crops are cultivated during spaceflight missions.

Jess M. Bunchek↗

Legume Organoleptic and Nutritional Analysis

Long-duration missions beyond low Earth orbit will encounter challenges in maintaining adequate nutrition and acceptability in the food system. In situ production of fresh produce can supplement nutrients deficient in the stored diet. Currently there is a limited number of crops that can be reliably grown for space crop production. Recent challenges with Veggie plant growth technical demonstrations (i.e. Tokyo Bekana Chinese cabbage interveinal chlorosis and necrosis when grown under elevated CO2 (~3000ppm) and narrow-band LED lighting) have highlighted the necessity to conduct rigorous ISS-relevant crop screening on the ground. Additionally, crops must be selected to address specific nutritional deficits as identified by HRP, with an emphasis on having a large diversity of crops available to meet nutritional requirements and crew acceptability. A variety of crop types are necessary to address known nutritional deficits in the stored astronaut diet, to include leafy greens (vitamin C, vitamin K, potassium), tomatoes and peppers (vitamin C, potassium, lycopene), and legumes (vitamin B1).Legumes are being evaluated for biological performance and suitability under ISS-like environmental conditions as well as nutritional content and acceptability. Legumes are one of the few pick and eat crop types that is an excellent source of Vitamin B1.In FY21-22 Biological and Physical Sciences (BPS) is funding screening of 32 legume crop cultivars at KSC, 16 of which will be down-selected based on horticultural performance and informal taste testing for further horticultural assessment and analysis. HRP in FY21-22 is funding organoleptic and nutritional analysis of these 16 down-selected legume cultivars to determine the most promising candidates for future space applications. Nutritional analysis will include full elemental, proximate (fat, protein, calories, carbohydrates, ash), and vitamins B1, C, K. This new work is currently postponed due to Covid-19.

L E Spencer↗

Production of Tuber-Inducing Factor

A process for making a substance that regulates the growth of potatoes and some other economically important plants has been developed. The process also yields an economically important by-product: potatoes. The particular growth-regulating substance, denoted tuber-inducing factor (TIF), is made naturally by, and acts naturally on, potato plants. The primary effects of TIF on potato plants are reducing the lengths of the main shoots, reducing the numbers of nodes on the main stems, reducing the total biomass, accelerating the initiation of potatoes, and increasing the edible fraction (potatoes) of the overall biomass. To some extent, these effects of TIF can override environmental effects that typically inhibit the formation of tubers. TIF can be used in the potato industry to reduce growth time and increase harvest efficiency. Other plants that have been observed to be affected by TIF include tomatoes, peppers, radishes, eggplants, marigolds, and morning glories. In the present process, potatoes are grown with their roots and stolons immersed in a nutrient solution in a recirculating hydroponic system. From time to time, a nutrient replenishment solution is added to the recirculating nutrient solution to maintain the required nutrient concentration, water is added to replace water lost from the recirculating solution through transpiration, and an acid or base is added, as needed, to maintain the recirculating solution at a desired pH level. The growing potato plants secrete TIF into the recirculating solution. The concentration of TIF in the solution gradually increases to a range in which the TIF regulates the growth of the plants.

Stutte, Gary W.↗

Genome sequence and characterization of a novel Pseudomonas putida phage, MiCath

Abstract Pseudomonads are ubiquitous bacteria with importance in medicine, soil, agriculture, and biomanufacturing. We report a novel Pseudomonas putida phage, MiCath, which is the first known phage infecting P. putida S12, a strain increasingly used as a synthetic biology chassis. MiCath was isolated from garden soil under a tomato plant using P. putida S12 as a host and was also found to infect four other P. putida strains. MiCath has a ~ 61 kbp double-stranded DNA genome which encodes 97 predicted open reading frames (ORFs); functions could only be predicted for 48 ORFs using comparative genomics. Functions include structural phage proteins, other common phage proteins (e.g., terminase), a queuosine gene cassette, a cas4 exonuclease, and an endosialidase. Restriction digestion analysis suggests the queuosine gene cassette encodes a pathway capable of modification of guanine residues. When compared to other phage genomes, MiCath shares at most 74% nucleotide identity over 2% of the genome with any sequenced phage. Overall, MiCath is a novel phage with no close relatives, encoding many unique gene products.

59 BASIC BIOLOGICAL SCIENCES↗

Physiological Response of Plants Grown on Porous Ceramic Tubes

This research involves the manipulation of the root-zone water potential for the purposes of discriminating the rate limiting step in the inorganic nutrient uptake mechanism utilized by higher plants. This reaction sequence includes the pathways controlled by the root-zone conditions such as water tension and gradient concentrations. Furthermore, plant based control mechanisms dictated by various protein productions are differentiated as well. For the nutrients limited by the environmental availability, the kinetics were modeled using convection and diffusion equations. Alternatively, for the nutrients dependent upon enzyme manipulations, the uptakes are modeled using Michaelis-Menten kinetics. In order to differentiate between these various mechanistic steps, an experimental apparatus known as the Porous Ceramic Tube - Nutrient Delivery System (PCT-NDS) was used. Manipulation of the applied suction pressure circulating a nutrient solution through this system imposes a change in the matric component of the water potential. This compensates for the different osmotic components of water potential dictated by nutrient concentration. By maintaining this control over the root-zone conditions, the rate limiting steps in the uptake of the essential nutrients into tomato plants (Lycopersicon esculentum cv. Cherry Elite) were differentiated. Results showed that the uptake of some nutrients were mass transfer limited while others were limited by the enzyme kinetics. Each of these were adequately modeled with calculations and discussions of the parameter estimations provided.

Tsao, David↗

Tomato growth as affected by root-zone temperature and the addition of gibberellic acid and kinetin to nutrient solutions

The effect of root-zone temperature on young tomato plants (Lycopersicon esculentum Mill. cv. Heinz 1350) was evaluated in controlled environments using a recirculating solution culture system. Growth rates were measured at root-zone temperatures of 15 degrees, 20 degrees, 25 degrees, and 30 degrees C in a near optimum foliar environment. Optimum growth occurred at 25 degrees to 30 degrees during the first 4 weeks of growth and 20 degrees to 25 degrees during the 5th and 6th weeks. Growth was severely restricted at 15 degrees. Four concentrations of gibberellic acid (GA3) and kinetin were added to the nutrient solution in a separate trial; root-zone temperature was maintained at 15 degrees and 25 degrees. Addition of 15 micromoles GA3 to solutions increased specific leaf area, total leaf area, and dry weight production of plants in both temperature treatments. GA3-induced growth stimulation was greater at 15 degrees than at 25 degrees. GA3 may promote growth by increasing leaf area, enhancing photosynthesis per unit leaf area, or both. Kinetic was not useful in promoting growth at either temperature.

NASA Program CELSS↗

Washing Away the Worries About Germs

Fresh fruits and vegetables have been in demand by orbiting astronauts since the early days of the Space Shuttle. As one can imagine, however, oranges, onions, tomatoes, garlic, and other fresh items can provide a cornucopia of smells in a closed environment such as the Space Shuttle or the International Space Station (ISS), especially when they begin to perish. It does not help that they are loaded onto the Space Shuttle up to 24 hours in advance of a launch, and that the on-orbit shelf life is just 2 to 3 days for most, due to a lack of refrigeration. While such produce adds significant variety to astronauts diets and increases their morale, the odor that emanates from it as it ages can cause nausea. One of the last things astronauts need is associating this healthy fare with feelings of nausea. NASA is currently investigating the use of a commercial sanitation product it helped develop with private industry to thoroughly cleanse and, thus, increase the shelf life of fruits and vegetables being sent into space. Meanwhile, this product is ripe for the picking for consumers looking to do the same, and more, on Earth.

Source record↗

Plant Growth Research for Future Food Production - Seed Microbiome Project

Humanity is currently on the precipice of a new era: one where human civilization is no longer bound to the confines of a single planet. Now, organizations like the National Aeronautics and Space Administration (NASA) have extended their areas of research and are beginning to focus not just on getting humans into space, but also to keep them safe, healthy, and sane. This focus falls under the Human Research Program (HRP). A focus of this program is "dedicated to discovering the best methods and technologies to support safe, productive human space travel" [1]. For "safe and productive space travel" to occur, astronauts must have proper nutrition [1]. While sending up large amounts of dried and packaged food with every shuttle might work fairly well in the short term, when only a few people need to be fed, it will not be sustainable, especially as NASA looks toward longer space journeys beyond the Earth's orbit. Research into this area falls under Advanced Life Support (ALS), whose mission is to develop regenerative life support systems to support future NASA long-duration missions [2]. This would involve growing crops in space to supplement astronaut diets [2]. An important, yet often overlooked, part of growing crops in any environment is the microbial organisms that inhabit the plants' microbiomes. The Seed Microbiome Project aims to investigate the microbial presence throughout the life stages of three crops, Mizuna Mustard, 'Outredgeous' Red Romaine Lettuce, and 'Red Robin' Tomato, that have either been or will be grown on the ISS.

Birtell, Eva↗

Preserving Perishables (Dormavac)

A new commercial product that can preserve perishable commodities for weeks without freezing, so that they can be shipped fresh without the cost of air freight, has been developed by Grumman Corporation, Bethpage, Long Island, New York. The development benefited from the company's experience in developing the environmental control system for the Lunar Module, which delivered Apollo astronauts to the surface of the moon. Called Dormavac, the system provides a commodity-preserving environment within an aluminum container that can be transported by truck, rail or ship. Dormavac creates a cold-but above freezing-environment with high relative humidity and very low air pressure. The saturated air minimizes commodity weight loss and the air is automatically changed several times an hour to flush away odors and harmful gases released by the commodities. According to company literature, Dormavac significantly extends the transportation life of perishables. For example, pork has a normal cold storage life of about seven days, beef two weeks and tomatoes three weeks; with Dormavac, pork remains fresh for three weeks, beef more than six weeks and tomatoes seven weeks or more. Dormavac is manufactured and marketed by Grumman Allied Industries, Woodbury, New York. In developing the system, Grumman Allied drew upon the technological resources of another company subsidiary, Grumman Aerospace. Engineers who had earlier worked on Lunar Module environmental control brought their know-how and experience to the Dormavac development.

Source record↗

Experiments Advance Gardening at Home and in Space

Aeroponics, the process of growing plants suspended in air without soil or media, provides clean, efficient, and rapid food production. Crops can be planted and harvested year-round without interruption, and without contamination from soil, pesticides, and residue. Aeroponic systems also reduce water usage by 98 percent, fertilizer usage by 60 percent, and eliminate pesticide usage altogether. Plants grown in aeroponic systems have been shown to absorb more minerals and vitamins, making the plants healthier and potentially more nutritious. The suspended system also has other advantages. Since the growing environment can be kept clean and sterile, the chances of spreading plant diseases and infections commonly found in soil and other growing media are greatly reduced. Also, seedlings do not stretch or wilt while their roots are forming, and once the roots are developed, the plants can be easily moved into any type of growing media without the risk of transplant shock. Lastly, plants tend to grow faster in a regulated aeroponic environment, and the subsequent ease of transplant to a natural medium means a higher annual crop yield. For example, tomatoes are traditionally started in pots and transplanted to the ground at least 28 days later; growers using an aeroponic system can transplant them just 10 days after starting the plants in the growing chamber. This accelerated cycle produces six tomato crops per year, rather than the traditional one to two crop cycles. These benefits, along with the great reduction in weight by eliminating soil and much of the water required for plant growth, illustrate why this technique has found such enthusiastic support from NASA. Successful long-term missions into deep space will require crews to grow some of their own food during flight. Aeroponic crops are also a potential source of fresh oxygen and clean drinking water, and every ounce of food produced and water conserved aboard a spacecraft reduces payload weight, decreasing launch costs and freeing room for other cargo.

Source record↗

Advanced Life Support Project: Crop Experiments at Kennedy Space Center

Crop production systems provide bioregenerative technologies to complement human crew life support requirements on long duration space missions. Kennedy Space Center has lead NASA's research on crop production systems that produce high value fresh foods, provide atmospheric regeneration, and perform water processing. As the emphasis on early missions to Mars has developed, our research focused on modular, scalable systems for transit missions, which can be developed into larger autonomous, bioregenerative systems for subsequent surface missions. Components of these scalable systems will include development of efficient light generating or collecting technologies, low mass plant growth chambers, and capability to operate in the high energy background radiation and reduced atmospheric pressures of space. These systems will be integrated with air, water, and thermal subsystems in an operational system. Extensive crop testing has been done for both staple and salad crops, but limited data is available on specific cultivar selection and breadboard testing to meet nominal Mars mission profiles of a 500-600 day surface mission. The recent research emphasis at Kennedy Space Center has shifted from staple crops, such as wheat, soybean and rice, toward short cycle salad crops such as lettuce, onion, radish, tomato, pepper, and strawberry. This paper will review the results of crop experiments to support the Exploration Initiative and the ongoing development of supporting technologies, and give an overview of capabilities of the newly opened Space Life Science (SLS) Lab at Kennedy Space Center. The 9662 square m (104,000 square ft) SLS Lab was built by the State of Florida and supports all NASA research that had been performed in Hanger-L. In addition to NASA research, the SLS Lab houses the Florida Space Research Institute (FSRI), responsible for co-managing the facility, and the University of Florida (UF) has established the Space Agriculture and Biotechnology Research and Education (SABRE) Center with several faculty.

Sager, John C.↗

Bioactive exometabolites drive maintenance competition in simple bacterial communities

During prolonged resource limitation, bacterial cells can persist in metabolically active states of non-growth. These maintenance periods, such as those experienced in stationary phase, can include upregulation of secondary metabolism and release of exometabolites into the local environment. As resource limitation is common in many environmental microbial habitats, we hypothesized that neighboring bacterial populations employ exometabolites to compete or cooperate during maintenance and that these exometabolite-facilitated interactions can drive community outcomes. Here, we evaluated the consequences of exometabolite interactions over the stationary phase among three environmental strains: Burkholderia thailandensis E264, Chromobacterium subtsugae ATCC 31532, and Pseudomonas syringae pv. tomato DC3000. We assembled them into synthetic communities that only permitted chemical interactions. We compared the responses (transcripts) and outputs (exometabolites) of each member with and without neighbors. We found that transcriptional dynamics were changed with different neighbors and that some of these changes were coordinated between members. The dominant competitor B. thailandensis consistently upregulated biosynthetic gene clusters to produce bioactive exometabolites for both exploitative and interference competition. These results demonstrate that competition strategies during maintenance can contribute to community-level outcomes. It also suggests that the traditional concept of defining competitiveness by growth outcomes may be narrow and that maintenance competition could be an additional or alternative measure.

59 BASIC BIOLOGICAL SCIENCES↗