Persistence of Escherichia coli in the microbiomes of red Romaine lettuce (Lactuca sativa cv. 'Outredgeous') and mizuna mustard (Brassica rapa var. japonica) - does seed sanitization matter?

BACKGROUND: Seed sanitization via chemical processes removes/reduces microbes from the external surfaces of the seed and thereby could have an impact on the plants' health or productivity. To determine the impact of seed sanitization on the plants' microbiome and pathogen persistence, sanitized and unsanitized seeds from two leafy green crops, red Romaine lettuce (Lactuca sativa cv. 'Outredgeous') and mizuna mustard (Brassica rapa var. japonica) were exposed to Escherichia coli and grown in controlled environment growth chambers simulating environmental conditions aboard the International Space Station. Plants were harvested at four intervals from 7 days post-germination to maturity. The bacterial communities of leaf and root were investigated using the 16S rRNA sequencing while quantitative polymerase chain reaction (qPCR) and heterotrophic plate counts were used to reveal the persistence of E. coli. RESULT: E. coli was detectable for longer periods of time in plants from sanitized versus unsanitized seeds and was identified in root tissue more frequently than in leaf tissue. 16S rRNA sequencing showed dynamic changes in the abundance of members of the phylum Proteobacteria, Firmicutes, and Bacteroidetes in leaf and root samples of both leafy crops. We observed minimal changes in the microbial diversity of lettuce or mizuna leaf tissue with time or between sanitized and unsanitized seeds. Beta-diversity showed that time had more of an influence on all samples versus the E. coli treatment. CONCLUSION: Our results indicated that the seed surface sanitization, a current requirement for sending seeds to space, could influence the microbiome. Insight into the changes in the crop microbiomes could lead to healthier plants and safer food supplementation.

Leaf yield and mineral content of mizuna in response to cut-and-come-again harvest, substrate particle size, and fertilizer formulation in a simulated spaceflight environment.

The Veggie plant-growth unit deployed onboard the International Space Station (ISS) grows leafy vegetables to supplement crew diets. "Cut-and-come-again" harvests are tested to maximize vegetative yield while minimizing crew time. Water, oxygen, and fertilizer delivery to roots of leafy greens growing in microgravity have become the center of attention for Veggie. Current Veggie technology wicks water into particulate root substrates incorporating controlled-release fertilizer (CRF). Mizuna mustard (Brassica rapa) was grown under ISS-like environments in ground-based Veggie-analogue units comparing crop response to combinations of two different substrate particle sizes, two different fertilizer formulations, and three leaf-harvest times from each plant. Fine-particle porous ceramic substrate (Profile©) was compared with a 40:60 mix of fine-particle porous ceramic Profile© + coarse porous ceramic Turface© substrate. Identical 18-6-8 (NPK) CRF formulations consisting of [50% fast-release (T70) + 50% intermediate-release (T100) prills] vs. [50% fast-release (T70) + 50% slow-release (T180) prills] were incorporated into each substrate, and leaf tissues were harvested from each treatment combination at 28, 48, and 56 days after sowing. The combination of T100 CRF in 100% Profile© substrate gave the highest fresh mass (FM) and leaf area (LA) across harvests, whereas T180 CRF in 40% Profile© gave the lowest. Dry-mass (DM) yields varied with effects on leaf area. Tissue nitrogen (N) and potassium (K) specific contents declined across harvests for all treatment combinations but tended to be highest for T100 CRF/100% Profile©, and lowest for T180 CRF/40% Profile©. These major macronutrients were taken up faster by roots growing in small particle-size substrate incorporating intermediate-rate CRF, but also were depleted faster from the same treatment combination, suggesting it may not continue to be the best combination for additional harvests. Micronutrients did not decline in tissue specific content across treatment combinations, but manganese (Mn) accumulated in leaf tissue across treatments and apparently comes mainly from the ceramic substrate, regardless of particle size. Substrate leachate analysis following final harvest indicated that pH remained in the range for nominal availability of mineral nutrients for root uptake, but electro-conductivity measurements suggested depletion of fertilizer salts from root zones, especially from the treatment combination supporting the highest yields and major macronutrient contents. Although 100% Profile© was the better growth substrate for mizuna in combination with intermediate-rate CRF and three cut-and-come-again harvests in ground-based studies, mixed-particle-size substrates may be a better choice for plant growth under microgravity conditions, where capillary forces predominant and tend to cause saturation of a fine medium with water. Since there were no statistically significant interactions between substrate and fertilizer in this study, our ground-based findings for CRF choice should translate to the best substrate choice for microgravity, but if NASA wants to consider additional cut-and-come-again harvests from the same mizuna plants, more complex CRF formulations likely will have to be investigated.

Microgreens for Home, Commercial, and Space Farming: A Comprehensive Update of the Most Recent Developments.

Microgreens are edible young plants that have recently attracted interest because of their color and flavor diversity, phytonutrient abundance, short growth cycle, and minimal space and nutrient requirements. They can be cultivated in a variety of systems from simple home gardens to sophisticated vertical farms with automated irrigation, fertilizer delivery, and lighting controls. Microgreens have also attracted attention from space agencies hoping that their sensory qualities can contribute to the diet of astronauts in microgravity and their cultivation might help maintain crew physical and psychological health on long-duration spaceflight missions. However, many technical challenges and data gaps for growing microgreensboth on and off Earth remain unaddressed. This review summarizes recent studies on multiple aspects of microgreens, including nutritional and socioeconomic benefits, cultivation systems, operative conditions, innovative treatments, autonomous facilities, and potential space applications. It also provides the authors' perspectives on the challenges to stimulating more extensive interdisciplinary research.

Comparison of two controlled-release fertilizer formulations for cut-and-come-again harvest yield and mineral content of Lactuca sativa L. cv. Outredgeous grown under International Space Station environmental conditions.

Red Romaine leaf lettuce (Lactuca sativa L. cv. Outredgeous) was grown in ground-based analogues of the Veggie plant-growth units used to grow salad vegetables for astronauts on the International Space Station (ISS). Plants were grown for 56 days with three "cut-and-come again" leaf harvests from the same plants. Six Biomass-Production-Systems-for-Education (BPSe) units were used to grow 'Outredgeous' ('OR') lettuce in a walk-in growth chamber under temperature, humidity, and LED-lighting conditions similar to those occurring in Veggie on ISS. Because of the ISS micro-gravity environment, both Veggie and ground-based BPSe units utilize one-way capillary wicking of water into an arcillite clay root substrate. In the present study, two different controlled-release fertilizer (CRF) formulations incorporated into the arcillite were compared for effects on 'OR' growth rate, overall yield, and mineral content of leaves harvested from the same plants 28, 48, and 56 days after planting. Both CRF treatments had a rapid-releasing T70 component that kept growth rate equivalent over the first two harvests. Growth rate for both CRF treatments increased from the first to the second harvest, but then declined from the second to the third harvest, more so for the slower-releasing T180 CRF than for the moderately-releasing T100 CRF. Tissue content of the macro-nutrients N, P, and K declined at each harvest for both CRFs, while content of the micro-nutrients B, Zn, and Mn increased. Although pH did not go out of the nominal range for availability of mineral nutrients to roots over the cropping cycle, and electrical-conductivity of rootzone salts was neither excessive nor depleted, tissue macronutrient depletion and micro-nutrient accumulation may have contributed to yield declines. Although either CRF formulation can support adequate yield of 'OR' lettuce over a 56-day period, the moderately-releasing T100 formulation tends to give slightly higher yield, especially during the last growth increment, and with non-deficient mineral content.

Feasibility of ultraviolet-light-emitting diodes as an alternative light source for photocatalysis.

The objective of this study was to determine whether ultraviolet-light-emitting diodes (UV-LEDs) could serve as an efficient photon source for heterogeneous photocatalytic oxidation (PCO). An LED module consisting of 12 high-power UV-A (lambda max = 365 nm) LEDs was designed to be interchangeable with a UV-A fluorescent black light blue (BLB) lamp for a bench scale annular reactor packed with silica-titania composite (STC) pellets. Lighting and thermal properties of the module were characterized to assess its uniformity and total irradiance. A forward current (I(F)) of 100 mA delivered an average irradiance of 4.0 mW cm(-2) at a distance of 8 mm, which is equivalent to the maximum output of the BLB, but the irradiance of the LED module was less uniform than that of the BLB. The LED and BLB reactors were tested for the oxidization of ethanol (50 ppm(v)) in a continuous-flow-through mode with 0.94 sec residence time. At the same average irradiance, the UV-A LED reactor resulted in a lower CO2 production rate (19.8 vs. 28.6 nmol L(-1) s(-1)), lower ethanol removal (80% vs. 91%), and lower mineralization efficiency (28% vs. 44%) than the UV-A BLB reactor. Ethanol mineralization was enhanced with the increase of the irradiance at the catalyst surface. This result suggests that reduced ethanol mineralization in the LED reactor relative to the BLB reactor at the same average irradiance could be attributed to the nonuniform irradiance over the photocatalyst, that is, a portion of the catalyst was exposed to less than the average irradiance. The potential of UV-A LEDs may be fully realized by optimizing the light distribution over the catalyst and utilizing their instantaneous "on" and "off" feature for periodic irradiation. Nevertheless, our results also showed that the current UV-A LED module had the same wall plug efficiency (WPE) of 13% as that of the UV-A BLB, demonstrating that UV-A LEDs are a viable photon source both in terms of WPE and PCO efficiency.

A Microbial Monitoring System Demonstrated on the International Space Station Provides a Successful Platform for Detection of Targeted Microorganisms.

Closed environments such as the International Space Station (ISS) and spacecraft for other planned interplanetary destinations require sustainable environmental control systems for manned spaceflight and habitation. These systems require monitoring for microbial contaminants and potential pathogens that could foul equipment or affect the health of the crew. Technological advances may help to facilitate this environmental monitoring, but many of the current advances do not function as expected in reduced gravity conditions. The microbial monitoring system (RAZOR® EX) is a compact, semi-quantitative rugged PCR instrument that was successfully tested on the ISS using station potable water. After a series of technical demonstrations between ISS and ground laboratories, it was determined that the instruments functioned comparably and provided a sample to answer flow in approximately 1 hour without enrichment or sample manipulation. Post-flight, additional advancements were accomplished at Kennedy Space Center, Merritt Island, FL, USA, to expand the instrument's detections of targeted microorganisms of concern such as water, food-borne, and surface microbes including Salmonella enterica serovar Typhimurium, Pseudomonas aeruginosa, Escherichia coli, and Aeromonas hydrophilia. Early detection of contaminants and bio-fouling microbes will increase crew safety and the ability to make appropriate operational decisions to minimize exposure to these contaminants.

Spatial Characterization of Microbial Communities on Multi-Species Leafy Greens Grown Simultaneously in the Vegetable Production Systems on the International Space Station.

The establishment of steady-state continuous crop production during long-term deep space missions is critical for providing consistent nutritional and psychological benefits for the crew, potentially improving their health and performance. Three technology demonstrations were completed achieving simultaneous multi-species plant growth and the concurrent use of two Veggie units on the International Space Station (ISS). Microbiological characterization using molecular and culture-based methods was performed on leaves and roots from two harvests of three leafy greens, red romaine lettuce (Lactuca sativa cv. 'Outredgeous'); mizuna mustard, (Brassica rapa var japonica); and green leaf lettuce, (Lactuca sativa cv. Waldmann's) and associated rooting pillow components and Veggie chamber surfaces. Culture based enumeration and pathogen screening indicated the leafy greens were safe for consumption. Surface samples of the Veggie facility and plant pillows revealed low counts of bacteria and fungi and are commonly isolated on ISS. Community analysis was completed with 16S rRNA amplicon sequencing. Comparisons between pillow components, and plant tissue types from VEG-03D, E, and F revealed higher diversity in roots and rooting substrate than the leaves and wick. This work provides valuable information for food production-related research on the ISS and the impact of the plant microbiome on this unique closed environment.

Growth and photosynthetic responses of Chinese cabbage (Brassica rapa L. cv. Tokyo Bekana) to continuously elevated carbon dioxide in a simulated Space Station "Veggie" crop-production environment.

Among candidate leafy vegetable species initially considered for astronauts to pick and eat from the Veggie plant-growth unit on the International Space Station (ISS), Chinese cabbage (Brassica rapa L. cv. Tokyo Bekana) ranked high in ground-based screening studies. However, subsequent attempts to optimize growth within rigorous ISS-like growth environments on the ground were frustrated by development of leaf chlorosis, necrosis, and uneven growth. 'Tokyo Bekana' ('TB') grown on ISS during the VEG-03B and C flights developed similar stress symptoms. After lengthy troubleshooting efforts to identify causes of sub-par growth in highly controlled environments, the super-elevated CO2 concentrations that plants on ISS are exposed to continuously (average of 2,800 µmol/mol) emerged as a candidate environmental condition responsible for the observed plant-stress symptoms. Subsequent ground-based studies found continuous exposure to ISS levels of CO2 under Veggie environmental and cultural conditions to significantly inhibit growth of 'TB' compared to near-Earth-normal CO2 controls. The present study investigated growth and gas-exchange responses of 'TB' to sub-ISS but still elevated CO2 levels (900 or 1,350 µmol/mol) in combination with other potential stressors related to ISS/Veggie compared to 450 µmol/mol CO2 controls. Shoot dry mass of plants grown at 450 µmol•mol-1 CO2 for 28 days was 96% and 80% higher than that of plants grown at 900 µmol•mol-1 CO2 and 1,350 µmol•mol-1 CO2, respectively. Leaf number and leaf area of controls were significantly higher than those of plants grown at 1,350 µmol•mol-1 CO2. Photosynthetic rate measured using a leaf cuvette was significantly lower for plants grown at 900 µmol•mol-1 CO2 than for controls. The ratio of leaf internal CO2 concentration (Ci) to cuvette ambient CO2 concentration (Ca) was significantly lower for plants grown at 450 µmol•mol-1 CO2 than for plants grown at elevated CO2. Thus, continuously elevated CO2 in combination with a Veggie cultivation system decreased growth, leaf area, and photosynthetic efficiency of Chinese cabbage 'Tokyo Bekana'. The results of this study suggest that 'Tokyo Bekana' is very sensitive to continuously elevated CO2 in such a growth environment, and indicate the need for improved environmental control of CO2 and possibly root-zone factors for successful crop production in the ISS spaceflight environment. Differential sensitivity of other salad crops to an ISS/Veggie growth environment also is possible, so it is important to mimic controllable ISS-like environmental conditions as precisely as possible during ground-based screening.

Microbiological and Nutritional Analysis of Lettuce Crops Grown on the International Space Station.

The ability to grow safe, fresh food to supplement packaged foods of astronauts in space has been an important goal for NASA. Food crops grown in space experience different environmental conditions than plants grown on Earth (e.g., reduced gravity, elevated radiation levels). To study the effects of space conditions, red romaine lettuce, Lactuca sativa cv 'Outredgeous,' plants were grown in Veggie plant growth chambers on the International Space Station (ISS) and compared with ground-grown plants. Multiple plantings were grown on ISS and harvested using either a single, final harvest, or sequential harvests in which several mature leaves were removed from the plants at weekly intervals. Ground controls were grown simultaneously with a 24-72 h delay using ISS environmental data. Food safety of the plants was determined by heterotrophic plate counts for bacteria and fungi, as well as isolate identification using samples taken from the leaves and roots. Molecular characterization was conducted using Next Generation Sequencing (NGS) to provide taxonomic composition and phylogenetic structure of the community. Leaves were also analyzed for elemental composition, as well as levels of phenolics, anthocyanins, and Oxygen Radical Absorbance Capacity (ORAC). Comparison of flight and ground tissues showed some differences in total counts for bacteria and yeast/molds (2.14 - 4.86 log10 CFU/g), while screening for select human pathogens yielded negative results. Bacterial and fungal isolate identification and community characterization indicated variation in the diversity of genera between leaf and root tissue with diversity being higher in root tissue, and included differences in the dominant genera. The only difference between ground and flight experiments was seen in the third experiment, VEG-03A, with significant differences in the genera from leaf tissue. Flight and ground tissue showed differences in Fe, K, Na, P, S, and Zn content and total phenolic levels, but no differences in anthocyanin and ORAC levels. This study indicated that leafy vegetable crops can produce safe, edible, fresh food to supplement to the astronauts' diet, and provide baseline data for continual operation of the Veggie plant growth units on ISS.

Potatoes as a Crop for Space Life Support: Effect of CO2, Irradiance, and Photoperiod on Leaf Photosynthesis and Stomatal Conductance.

Potatoes (Solanum tuberosum L.) have been suggested as a candidate crop for future space missions, based on their high yields of nutritious tubers and high harvest index. Three cultivars of potato, cvs. Norland, Russet Burbank, and Denali were grown in walk-in growth rooms at 400 and 800 µmol m-2 s-1 photosynthetic photon flux (PPF), 12-h L/12-h D and 24-h L/0 h D photoperiods, and 350 and 1,000 ppm [CO2]. Net photosynthetic rates (Pnet) and stomatal conductance (gs) of upper canopy leaves were measured at weekly intervals from 3 through 12 weeks after planting. Increased PPF resulted in increased Pnet rates at both [CO2] levels and both photoperiods, but the effect was most pronounced under the 12-h photoperiod. Increased [CO2] increased Pnet for both PPFs under the 12-h photoperiod, but decreased Pnet under the 24-h photoperiod. Increased PPF increased gs for both [CO2] levels and both photoperiods. Increased [CO2] decreased gs for both PPFs for the 12-h photoperiod, but caused only a slight decrease under the 24-h photoperiod. Leaf Pnet rates were highest with high PPF (800), elevated [CO2] (1,000), and a 12-h photoperiod, while growing the plants under continuous (24-h) light resulted in lower leaf photosynthetic rates for all combinations of PPF and [CO2]. The responses of leaf photosynthetic rates are generally consistent with prior published data on the plant biomass from these same studies (Wheeler et al., Crop Sci. 1991) and suggest that giving more light with a 24-h photoperiod can increase biomass in some cases, but the leaf Pnet and overall photosynthetic efficiency drops.

Influence of Microgravity Environment on Root Growth, Soluble Sugars, and Starch Concentration of Sweetpotato Stem Cuttings.

Because sweetpotato [Ipomoea batatas (L.) Lam.] stem cuttings regenerate very easily and quickly, a study of their early growth and development in microgravity could be useful to an understanding of morphological changes that might occur under such conditions for crops that are propagated vegetatively. An experiment was conducted aboard a U.S. Space Shuttle to investigate the impact of microgravity on root growth, distribution of amyloplasts in the root cells, and on the concentration of soluble sugars and starch in the stems of sweetpotatoes. Twelve stem cuttings of 'Whatley/Loretan' sweetpotato (5 cm long) with three to four nodes were grown in each of two plant growth units filled with a nutrient agarose medium impregnated with a half-strength Hoagland solution. One plant growth unit was flown on Space Shuttle Colombia for 5 days, whereas the other remained on the ground as a control. The cuttings were received within 2 h postflight and, along with ground controls, processed in approximately 45 min. Adventitious roots were counted, measured, and fixed for electron microscopy and stems frozen for starch and sugar assays. Air samples were collected from the headspace of each plant growth unit for postflight determination of carbon dioxide, oxygen, and ethylene levels. All stem cuttings produced adventitious roots and growth was quite vigorous in both ground-based and flight samples and, except for a slight browning of some root tips in the flight samples, all stem cuttings appeared normal. The roots on the flight cuttings tended to grow in random directions. Also, stem cuttings grown in microgravity had more roots and greater total root length than ground-based controls. Amyloplasts in root cap cells of ground-based controls were evenly sedimented toward one end compared with a more random distribution in the flight samples. The concentration of soluble sugars, glucose, fructose, and sucrose and total starch concentration were all substantially greater in the stems of flight samples than those found in the ground-based samples. Carbon dioxide levels were 50% greater and oxygen marginally lower in the flight plants, whereas ethylene levels were similar and averaged less than 10 nL.L (-1). Despite the greater accumulation of carbohydrates in the stems, and greater root growth in the flight cuttings, overall results showed minimal differences in cell development between space flight and ground-based tissues. This suggests that the space flight environment did not adversely impact sweetpotato metabolism and that vegetative cuttings should be an acceptable approach for propagating sweetpotato plants for space applications.

Exposure of Arabidopsis thaliana to hypobaric environments: implications for low-pressure bioregenerative life support systems for human exploration missions and terraforming on Mars.

Understanding how hypobaria can affect net photosynthetic (P (net)) and net evapotranspiration rates of plants is important for the Mars Exploration Program because low-pressured environments may be used to reduce the equivalent system mass of near-term plant biology experiments on landers or future bioregenerative advanced life support systems. Furthermore, introductions of plants to the surface of a partially terraformed Mars will be constrained by the limits of sustainable growth and reproduction of plants to hypobaric conditions. To explore the effects of hypobaria on plant physiology, a low-pressure growth chamber (LPGC) was constructed that maintained hypobaric environments capable of supporting short-term plant physiological studies. Experiments were conducted on Arabidopsis thaliana maintained in the LPGC with total atmospheric pressures set at 101 (Earth sea-level control), 75, 50, 25 or 10 kPa. Plants were grown in a separate incubator at 101 kPa for 6 weeks, transferred to the LPGC, and acclimated to low-pressure atmospheres for either 1 or 16 h. After 1 or 16 h of acclimation, CO(2) levels were allowed to drawdown from 0.1 kPa to CO(2) compensation points to assess P (net) rates under different hypobaric conditions. Results showed that P (net) increased as the pressures decreased from 101 to 10 kPa when CO(2) partial pressure (pp) values were below 0.04 kPa (i.e., when ppCO2 was considered limiting). In contrast, when ppCO(2) was in the nonlimiting range from 0.10 to 0.07 kPa, the P (net) rates were insensitive to decreasing pressures. Thus, if CO(2 )concentrations can be kept elevated in hypobaric plant growth modules or on the surface of a partially terraformed Mars, P (net) rates may be relatively unaffected by hypobaria. Results support the conclusions that (i) hypobaric plant growth modules might be operated around 10 kPa without undue inhibition of photosynthesis and (ii) terraforming efforts on Mars might require a surface pressure of at least 10 kPa (100 mb) for normal growth of deployed plant species.

Light-emitting diodes as an illumination source for plants: a review of research at Kennedy Space Center.

The provision of sufficient light is a fundamental requirement to support long-term plant growth in space. Several types of electric lamps have been tested to provide radiant energy for plants in this regard, including fluorescent, high-pressure sodium, and metal halide lamps. These lamps vary in terms of spectral quality, which can result in differences in plant growth and morphology. Current lighting research for space-based plant culture is focused on innovative lighting technologies that demonstrate high electrical efficiency and reduced mass and volume. Among the lighting technologies considered for space are light-emitting diodes (LEDs). The combination of red and blue LEDs has proven to be an effective lighting source for several crops, yet the appearance of plants under red and blue lighting is purplish gray, making visual assessment of plant health difficult. Additional green light would make the plant leaves appear green and normal, similar to a natural setting under white light, and may also offer psychological benefits for the crew. The addition of 24% green light (500-600 nm) to red and blue LEDs enhanced the growth of lettuce plants compared with plants grown under cool white fluorescent lamps. Coincidentally, these plants grown under additional green light would have the additional aesthetic appeal of a green appearance.

Carbon balance in bioregenerative life support systems: some effects of system closure, waste management, and crop harvest index.

In Advanced Life Support (ALS) systems with bioregenerative components, plant photosynthesis would be used to produce O2 and food, while removing CO2. Much of the plant biomass would be inedible and hence must be considered in waste management. This waste could be oxidized (e.g., incinerated or aerobically digested) to resupply CO2 to the plants, but this would not be needed unless the system were highly closed with regard to food. For example, in a partially closed system where some of the food is grown and some is imported, CO2 from oxidized waste when combined with crew and microbial respiration could exceed the CO2 removal capability of the plants. Moreover, it would consume some O2 produced from photosynthesis that could have been used by the crew. For partially closed systems it would be more appropriate to store or find other uses for the inedible biomass and excess carbon, such as generating soils or growing woody plants (e.g., dwarf fruit trees). Regardless of system closure, high harvest crops (i.e., crops with a high edible to total biomass ratio) would increase food production per unit area and O2 yields for systems where waste biomass is oxidized to recycle CO2. Such interlinking effects between the plants and waste treatment strategies point out the importance of oxidizing only that amount of waste needed to optimize system performance.

Influence of nitrogen nutrition management on biomass partitioning and nitrogen use efficiency indices in hydroponically grown potato.

The National Aeronautics and Space Administration (NASA) has been conducting controlled environment research with potatoes (Solanum tuberosum L.) in recirculating nutrient film technique (NFT)-hydroponic systems as a human life support component during long-duration spaceflight. Standard nutrient solution management approaches include constant pH regulation with nitric acid (HNO3) and daily adjustment of electrical conductivity (EC) equivalent to half-strength modified Hoagland's solution, where nitrate (NO3-) is the sole nitrogen (N) source. Although tuber yields have been excellent with such an approach, N use efficiency indices are expected to be low relative to tuber biomass production. Furthermore, the high amount of N used in NFT-hydroponics, typically results in high inedible biomass, which conflicts with the need to minimize system mass, volume, and expenditure of resources for long-duration missions. More effective strategies of N fertilization need to be developed to more closely match N supply with demand of the crop. Hence, the primary objective of this study was to identify the optimal N management regime and plant N requirement to achieve high yields and to avoid inefficient use of N and excess inedible biomass production. In separate 84-day cropping experiments, three N management protocols were tested. Treatments which decreased NO3(-)-N supply indirectly through lowering nutrient solution EC (Expt. I), or disabling pH control, and/or supplying NH4(+)-N (Expt. III) did not significantly benefit tuber yield, but did influence N use efficiency indices. When supplied with an external 7.5 mM NO3(-)-N for the first 42 days after planting (DAP), lowered to 1.0 mM NO3(-)-N during the final 42 days (Expt. II), plants were able to achieve yields on par with plants which received constant 7.5 mM NO3(-)-N (control). By abruptly decreasing N supply at tuber initiation in Expt. II, less N was taken up and accumulated by plants compared to those which received high constant N (control). However, proportionately more plant accumulated N was used (N use efficiency) to produce tuber biomass when N supply was abruptly lowered at tuber initiation in Expt. II. Hence, a hydroponic nutrient solution N management system may be modified to elicit greater plant N-use while maintaining overall high tuber yield as opposed to achieving high tuber yields through excess N supply and shoot growth.

Ethylene production throughout growth and development of plants.

Ethylene production by 10 or 20 m2 stands of wheat, soybean, lettuce, potato, and tomato was monitored throughout growth and development in an atmospherically closed plant chamber. Chamber ethylene levels varied among species and rose during periods of canopy expansion and rapid growth for all species. Following this, ethylene levels either declined during seed fill and maturation for wheat and soybean, or remained relatively constant for potato and tomato (during flowering and early fruit development). Lettuce plants were harvested during rapid growth and peak ethylene production. Chamber ethylene levels increased rapidly during tomato ripening, reaching concentrations about 10 times that measured during vegetative growth. The highest ethylene production rates during vegetative growth ranged from 1.6 to 2.5 nmol m-2 d-1 during rapid growth of lettuce and wheat stands, or about 0.3 to 0.5 nmol g-1 fresh weight per hour. Estimates of stand ethylene production during tomato ripening showed that rates reached 43 nmol m-2 d-1 in one study and 93 nmol m-2 d-1 in a second study with higher lighting, or about 50x that of the rate during vegetative growth of tomato. In a related test with potato, the photoperiod was extended from 12 to 24 hours (continuous light) at 58 days after planting (to increase tuber yield), but this change in the environment caused a sharp increase in ethylene production from the basal rate of 0.4 to 6.2 nmol m-2 d-1. Following this, the photoperiod was changed back to 12 h at 61 days and ethylene levels decreased. The results suggest three separate categories of ethylene production were observed with whole stands of plants: 1) production during rapid vegetative growth, 2) production during climacteric fruit ripening, and 3) production from environmental stress.

Green-light supplementation for enhanced lettuce growth under red- and blue-light-emitting diodes.

Plants will be an important component of future long-term space missions. Lighting systems for growing plants will need to be lightweight, reliable, and durable, and light-emitting diodes (LEDs) have these characteristics. Previous studies demonstrated that the combination of red and blue light was an effective light source for several crops. Yet the appearance of plants under red and blue lighting is purplish gray making visual assessment of any problems difficult. The addition of green light would make the plant leave appear green and normal similar to a natural setting under white light and may also offer a psychological benefit to the crew. Green supplemental lighting could also offer benefits, since green light can better penetrate the plant canopy and potentially increase plant growth by increasing photosynthesis from the leaves in the lower canopy. In this study, four light sources were tested: 1) red and blue LEDs (RB), 2) red and blue LEDs with green fluorescent lamps (RGB), 3) green fluorescent lamps (GF), and 4) cool-white fluorescent lamps (CWF), that provided 0%, 24%, 86%, and 51% of the total PPF in the green region of the spectrum, respectively. The addition of 24% green light (500 to 600 nm) to red and blue LEDs (RGB treatment) enhanced plant growth. The RGB treatment plants produced more biomass than the plants grown under the cool-white fluorescent lamps (CWF treatment), a commonly tested light source used as a broad-spectrum control.

Water cycle and its management for plant habitats at reduced pressures.

Experimental and mathematical models were developed for describing and testing temperature and humidity parameters for plant production in bioregenerative life support systems. A factor was included for analyzing systems operating at low (10-101.3 kPa) pressure to reduce gas leakage and structural mass (e.g., inflatable greenhouses for space application). The expected close relationship between temperature and relative humidity was observed, along with the importance of heat exchanger coil temperature and air circulation rate. The presence of plants in closed habitats results in increased water flux through the system. Changes in pressure affect gas diffusion rates and surface boundary layers, and change convective transfer capabilities and water evaporation rates. A consistent observation from studies with plants at reduced pressures is increased evapotranspiration rates, even at constant vapor pressure deficits. This suggests that plant water status is a critical factor for managing low-pressure production systems. The approach suggested should help space mission planners design artificial environments in closed habitats.

Bioavailability of iron from spinach using an in vitro/human Caco-2 cell bioassay model.

Spinach (Spinacia oleracea) cv Whitney was tested for iron bioavailabilty using an in vitro human intestinal cell culture ferritin bioassay technique previously developed. Spinach was cultured in a growth chamber for 33 days, harvested, and freeze-dried. Total iron in the samples was an average of 71 micrograms/g dry weight. Spinach was digested in vitro (pepsin and 0.1 M HCl followed by pancreatin and 0.1 M NaHCO3) with and without the addition of supplemental ascorbic acid. Caco-2 cell cultures were used to determine iron bioavailability from the spinach mixtures. Production of the iron-binding protein ferritin in the Caco-2 cells showed the supplemental ascorbic acid doubled bioavailability of iron from spinach. The data show fresh spinach is a poor source of iron, and emphasize the importance of evaluation of whole meals rather than single food items. The data support the usefulness of the in vitro/Caco-2 cell ferritin bioassay model for prescreening of space flight diets for bioavailable iron.

Host-microbe interactions in microgravity: assessment and implications.

Spaceflight imposes several unique stresses on biological life that together can have a profound impact on the homeostasis between eukaryotes and their associated microbes. One such stressor, microgravity, has been shown to alter host-microbe interactions at the genetic and physiological levels. Recent sequencing of the microbiomes associated with plants and animals have shown that these interactions are essential for maintaining host health through the regulation of several metabolic and immune responses. Disruptions to various environmental parameters or community characteristics may impact the resiliency of the microbiome, thus potentially driving host-microbe associations towards disease. In this review, we discuss our current understanding of host-microbe interactions in microgravity and assess the impact of this unique environmental stress on the normal physiological and genetic responses of both pathogenic and mutualistic associations. As humans move beyond our biosphere and undergo longer duration space flights, it will be essential to more fully understand microbial fitness in microgravity conditions in order to maintain a healthy homeostasis between humans, plants and their respective microbiomes.