Substrate Matters: Ionic Silver Alters Lettuce Growth, Nutrient Uptake, and Root Microbiome in a Hydroponics System.

Ionic silver (Ag+) is being investigated as a residual biocide for use in NASA spacecraft potable water systems on future crewed missions. This water will be used to irrigate future spaceflight crop production systems. We have evaluated the impact of three concentrations (31 ppb, 125 ppb, and 500 ppb) of ionic silver biocide solutions on lettuce in an arcillite (calcinated clay particle substrate) and hydroponic (substrate-less) growth setup after 28 days. Lettuce plant growth was reduced in the hydroponic samples treated with 31 ppb silver and severely stunted for samples treated at 125 ppb and 500 ppb silver. No growth defects were observed in arcillite-grown lettuce. Silver was detectable in the hydroponic-grown lettuce leaves at each concentration but was not detected in the arcillite-grown lettuce leaves. Specifically, when 125 ppb silver water was applied to a hydroponics tray, Ag+ was detected at an average amount of 7 µg/g (dry weight) in lettuce leaves. The increase in Ag+ corresponded with a decrease in several essential elements in the lettuce tissue (Ca, K, P, S). In the arcillite growth setup, silver did not impact the plant root zone microbiome in terms of alpha diversity and relative abundance between treatments and control. However, with increasing silver concentration, the alpha diversity increased in lettuce root samples and in the water from the hydroponics tray samples. The genera in the hydroponic root and water samples were similar across the silver concentrations but displayed different relative abundances. This suggests that ionic silver was acting as a selective pressure for the microbes that colonize the hydroponic water. The surviving microbes likely utilized exudates from the stunted plant roots as a carbon source. Analysis of the root-associated microbiomes in response to silver showed enrichment of metagenomic pathways associated with alternate carbon source utilization, fatty-acid synthesis, and the ppGpp (guanosine 3'-diphosphate 5'-diphosphate) stringent response global regulatory system that operates under conditions of environmental stress. Nutrient solutions containing Ag+ in concentrations greater than 31 ppb in hydroponic systems lacking cation-exchange capacity can severely impact crop production due to stunting of plant growth.

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.

Crew time in a space greenhouse using data from analog missions and Veggie.

Crew time requirements for human space exploration missions is as critical as mass, energy, and volume requirements. However, it has only been sporadically recorded in past analog and space missions for plant cultivation. In this retrospective study on crew time data collected in various analog facilities and on the Veggie hardware on ISS, we propose a methodology for efficient categorizing and reporting of crew time in space plant growth systems. Crew time is difficult to capture in operational environments, and this study intends to harmonize these efforts among different locations. This article also provides a current database for required crew time in several plant growth hardware and facilities, on the ISS, and on Earth. These data could serve mission planners as a baseline to establish standardized activities and extrapolate crew time needed to operate future plant growth units. Finally, we discuss how crew time needed for plant cultivation will change in future exploration missions, based on choices made for plant species, watering systems, level of automation, and use of virtual assistants, among others. Crew time will need to be accounted for as a decisive factor to design future space greenhouse modules.

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.

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.

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.

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.

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.

Early seedling response of six candidate crop species to increasing levels of blue light.

Light emitting diode (LED) lighting technology for crop production is advancing at a rapid pace, both in terms of the technology itself (e.g., spectral composition and efficiency), and the research that the technological advances have enabled. The application of LED technology for crop production was first explored as a tool for improving the safety and reliability of plant-based bioregenerative life-support systems for long duration human space exploration. Developing and optimizing the lighting environment (spectral quality and quantity) for bioregenerative life-support applications and other controlled environment plant production applications, such as microgreens and sprout production, continues to be an active area of research and LED technology development. This study examines the influence of monochromatic and dichromatic red and blue light on the early development of six food crop species; Cucumis sativa, Solanum lycopersicum, Glycine max, Raphanus sativus, Pisum sativum, and Capsicum annum. Results support previous findings that light responses are often species specific. The results also support the assertion that monochromatic light can interfere with the normal interaction of various photoreceptors (co-action disruption) resulting in intermediate and sometimes unpredictable responses to a given light environment. The nature of the responses reported inform both bioregenerative life-support designs as well as light quality selection for the production of controlled environment crops.

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.

Root restriction: A tool for improving volume utilization efficiency in bioregenerative life-support systems.

The objective of this study was to evaluate root restriction as a tool to increase volume utilization efficiency in spaceflight crop production systems. Bell pepper plants (Capsicum annuum cv. California Wonder) were grown under restricted rooting volume conditions in controlled environment chambers. The rooting volume was restricted to 500ml and 60ml in a preliminary trial, and 1500ml (large), 500ml (medium), and 250ml (small) for a full fruiting trial. To reduce the possible confounding effects of water and nutrient restrictions, care was taken to ensure an even and consistent soil moisture throughout the study, with plants being watered/fertilized several times daily with a low concentration soluble fertilizer solution. Root restriction resulted in a general reduction in biomass production, height, leaf area, and transpiration rate; however, the fruit production was not significantly reduced in the root restricted plants under the employed environmental and horticultural conditions. There was a 21% reduction in total height and a 23% reduction in overall crown diameter between the large and small pot size in the fruiting study. Data from the fruiting trial were used to estimate potential volume utilization efficiency improvements for edible biomass in a fixed production volume. For fixed lighting and rooting hardware situations, the majority of improvement from root restriction was in the reduction of canopy area per plant, while height reductions could also improve volume utilization efficiency in high stacked or vertical agricultural systems.

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.

Genome-wide expression analysis of reactive oxygen species gene network in Mizuna plants grown in long-term spaceflight.

BACKGROUND: Spaceflight environment have been shown to generate reactive oxygen species (ROS) and induce oxidative stress in plants, but little is known about the gene expression of the ROS gene network in plants grown in long-term spaceflight. The molecular response and adaptation to the spaceflight environment of Mizuna plants harvested after 27 days of cultivation onboard the International Space Station (ISS) were measured using genome-wide mRNA expression analysis (mRNA-Seq). RESULTS: Total reads of transcripts from the Mizuna grown in the ISS as well as on the ground by mRNA-Seq showed 8,258 and 14,170 transcripts up-regulated and down-regulated, respectively, in the space-grown Mizuna when compared with those from the ground-grown Mizuna. A total of 20 in 32 ROS oxidative marker genes were up-regulated, including high expression of four hallmarks, and preferentially expressed genes associated with ROS-scavenging including thioredoxin, glutaredoxin, and alternative oxidase genes. In the transcription factors of the ROS gene network, MEKK1-MKK4-MPK3, OXI1-MKK4-MPK3, and OXI1-MPK3 of MAP cascades, induction of WRKY22 by MEKK1-MKK4-MPK3 cascade, induction of WRKY25 and repression of Zat7 by Zat12 were suggested. RbohD and RbohF genes were up-regulated preferentially in NADPH oxidase genes, which produce ROS. CONCLUSIONS: This large-scale transcriptome analysis revealed that the spaceflight environment induced oxidative stress and the ROS gene network activation in the space-grown Mizuna. Among transcripts altered in expression by space conditions, some were common genes response to abiotic and biotic stress. Furthermore, certain genes were exclusively up-regulated in Mizuna grown on the ISS. Surprisingly, Mizuna grew in space normally, as well as on the ground, demonstrating that plants can acclimate to long-term exposure in the spaceflight environment by reprogramming the expression of the ROS gene network.

Transcriptional and metabolic insights into the differential physiological responses of arabidopsis to optimal and supraoptimal atmospheric CO2.

BACKGROUND: In tightly closed human habitats such as space stations, locations near volcano vents and closed culture vessels, atmospheric CO(2) concentration may be 10 to 20 times greater than Earth's current ambient levels. It is known that super-elevated (SE) CO(2) (>1,200 µmol mol(-1)) induces physiological responses different from that of moderately elevated CO(2) (up to 1,200 µmol mol(-1)), but little is known about the molecular responses of plants to supra-optimal [CO(2)]. METHODOLOGY/PRINCIPAL FINDINGS: To understand the underlying molecular causes for differential physiological responses, metabolite and transcript profiles were analyzed in aerial tissue of Arabidopsis plants, which were grown under ambient atmospheric CO(2) (400 µmol mol(-1)), elevated CO(2) (1,200 µmol mol(-1)) and SE CO(2) (4,000 µmol mol(-1)), at two developmental stages early and late vegetative stage. Transcript and metabolite profiling revealed very different responses to elevated versus SE [CO(2)]. The transcript profiles of SE CO(2) treated plants were closer to that of the control. Development stage had a clear effect on plant molecular response to elevated and SE [CO(2)]. Photosynthetic acclimation in terms of down-regulation of photosynthetic gene expression was observed in response to elevated [CO(2)], but not that of SE [CO(2)] providing the first molecular evidence that there appears to be a fundamental disparity in the way plants respond to elevated and SE [CO(2)]. Although starch accumulation was induced by both elevated and SE [CO(2)], the increase was less at the late vegetative stage and accompanied by higher soluble sugar content suggesting an increased starch breakdown to meet sink strength resulting from the rapid growth demand. Furthermore, many of the elevated and SE CO(2)-responsive genes found in the present study are also regulated by plant hormone and stress. CONCLUSIONS/SIGNIFICANCE: This study provides new insights into plant acclimation to elevated and SE [CO(2)] during development and how this relates to stress, sugar and hormone signaling.

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.

Super-elevated CO2 interferes with stomatal response to ABA and night closure in soybean (Glycine max).

Studies have shown stomatal conductance (g(s)) of plants exposed to super-elevated CO2 (>5000micromol mol(-1)) increases in several species, in contrast to a decrease of g(s) caused by moderate CO2 enrichment. We conducted a series of experiments to determine whether super-elevated CO2 alters stomatal development and/or interferes with stomatal closure in soybean (Glycine max). Plants were grown at nominal ambient (400), elevated (1200) and super-elevated (10,000micromol mol(-1)) CO2 in controlled environmental chambers. Stomatal density of the plant leaf was examined by a scanning electron microscope (SEM), while the stomatal response to the application of exogenous abscisic acid (ABA), a phytohormone associated with water stress and stomatal control, was investigated in intact growing plants by measuring the g(s) of abaxial leaf surfaces using a steady-state porometer. Relative to the control (400micromol mol(-1) CO2) plants, daytime stomatal conductance (g(s,day)) of the plants grown under 1200 and 10,000micromol mol(-1) CO2 was reduced by 38% and 15%, respectively. Dark period stomatal conductance (g(s,night)) was unaffected by growing under 1200mumol mol(-1) CO2) but dramatically increased under 10,000micromol mol(-1) CO2. Stomatal density increased by 10% in the leaves of 10,000micromol mol(-1) CO2-grown plants, which in part contributed to the higher g(s,night) values. Elevating [CO2] to 1200micromol mol(-1) enhanced ABA-induced stomatal closure, but further increasing CO2 to 10,000micromol mol(-1) significantly reduced ABA-induced stomatal closure. These results demonstrated that stomatal response to ABA is CO2 dependent. Hence, a stomatal failure to effectively respond to an ABA signal and to close at night under extremely high CO2 may increase plants susceptibility to other abiotic stresses.

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.