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N.C. Yorio, C.L. Mackowiak, R.M. Wheeler, and G.W. Stutte

The effects of elevated CO2 on stomatal density and index were investigated for five crop species currently being studied for NASA's Advanced Life Support program. Lettuce (cv. Waldmann's Green) and radish (cv. Giant White Globe) were grown at 400, 1000, 5000, or 10,000 μmol·mol–1 CO2, tomato (cvs. Red Robin and Reimann Philip 75/59) were grown at 400, 1200, 5000, or 10,000 μmol·mol–1 CO2, and wheat (cv. Yecora Rojo) and potato (cv. Denali) were grown at 400, 1000, or 10,000 μmol·mol–1 CO2 within controlled-environment growth chambers using nutrient film technique hydroponics. Leaf impressions were made by applying clear silicone-based RTV coating to the adaxial and abaxial leaf surfaces of three canopy leaves of each crop at each CO2 treatment. Impressions were examined using a light microscope, whereby the number of stomatal complexes and epidermal cells were counted to calculate stomatal density and stomatal index. Results indicate that stomatal density increased for lettuce and radish at 10,000 μmol·mol–1 CO2, whereas tomato density was highest at 1200 μmol·mol–1 CO2. Potato had the lowest density at 1000 μmol·mol–1 CO2, and there was no effect of CO2 on density for wheat. Stomatal index correlated with density for lettuce and tomato; however, stomatal index for radish, potato, and wheat was not influenced by CO2. This suggests that there may be a species-specific CO2 response to epidermal cell size that influences stomatal density and stomatal index.

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G.W. Stutte, N.C. Yorio, C.L. Mackowiak, and R.M. Wheeler

This experiment was performed to test the hypothesis that tuber formation in potato is inhibited by short-term increases in root-zone temperature. Micro-propagated potato cv. Norland plantlets were grown in recirculating nutrient film culture under daylight fluorescent lamps at 350 μmol·m–2·s–1 PPF with at 20/16°C thermocycle at 1200 μmol·mol–1 CO2 under inductive (12-hr light/12-hr dark) or non-inductive (12-hr light/12-hr dark with a 15-min light break 6 hr into the cycle) photoperiods for 42 days. Root-zone treatments consisted of continuous 18°C, continuous 24°C, 18°C with a 24°C cycle between 14 and 21 DAP (prior to tuber initiation), and 18°C with a 24°C cycle between 21 and 28 DAP (during the period of tuber initiation). The root-zone temperature was maintained with a recirculating, temperature-controlled, heat-exchange coil submerged in each nutrient solution. Warm root-zone temperatures did not inhibit tuber formation under an inductive photoperiod. The non-inductive photoperiod resulted in a 65% reduction in tuber biomass compared to the inductive photoperiod. Continuous 24°C and exposure to 24°C prior to tuber initiation reduced tuber formation an additional 40% under the non-inductive photoperiod. Both continuous and transient 24°C root-zone temperatures increased biomass partitioning to root/stolons compared to the 18°C treatment under both photoperiods. Total plant biomass was highest in plants exposed to continuous 24°C under both photoperiods. Results suggest that transient episodes of warm (24°C) root-zone temperature do not inhibit tuber formation in potato under inductive photoperiods. However, transient episodes of warm (24°C) root-zone temperatures did interact with stage of development under the non-inductive photoperiod.

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R.M. Wheeler, G.W. Stutte, C.L. Mackowiak, N.C. Yorio, and L.M. Ruffe

Potatoes (Solanum tuberosum L.) have been grown successfully with a recirculating nutrient film technique (NFT) when a fresh nutrient solution is used for each planting. During the past year, we conducted two studies in which the same nutrient solution was used for successive plantings (EC and pH were maintained at 0.12 S·m–1 and 5.8). Results showed that successive plantings became prematurely induced (tubers initiating near 20 days after planting–DAP), causing stunted shoot growth and reduced yields per plant. When “old” nutrient solution from a continuous production system was regularly added to a newly started plant system maintained under a non-inductive environment (12-h photoperiod with night break of 6 h into dark), tubers formed on “old” nutrient solution plants (24 DAP), but not on “new” solution plants. When charcoal water filters were placed on the systems, plants grown on either “old” or “new” nutrient solutions showed no tuber initiation (plants harvested at 42 DAP). Results suggest that a tuber-inducing factor(s) emanating from the plants accumulates in the nutrient solution over time and that the factor(s) can be removed by charcoal absorption.

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C.L. Mackowiak, G.W. Stutte, R.M. Wheeler, and N.C. Yorio

The growth of candidate crops in high CO2 environments is being investigated as part of NASA's goal of using higher plants for bioregenerative life support systems. Tomato (Lycopersicon esculentum Mill.) cvs. Red Robin and Reimann Philipp were grown in recirculating hydroponics at 400, 1200, 5000, or 10,000 μmol·mol–1 CO2 for 105 days. The plants received a 12/12 hour photo-period at 500 μmol·m–2·s–1 PPF, 26/22°C (light/dark), and 65% continuous relative humidity. Stomatal conductance increased at the highest CO2 levels, which is similar to what we have reported with Soybean, radish, and potato. Fruit number increased with increasing CO2, where Red Robin produced 663 fruit/m2 and Reimann Philipp produced 6870 fruit/m2 at 10,000 μmol·mol–1 CO2. Fruit fresh mass was greatest at 10,000 μmol·mol–1 CO2 for Red Robin (7.4 kg·m–2) and at 5000 μmol·mol–1 CO2 for Reimann Philipp (27 kg·m–2), suggesting that very high CO2 was not detrimental to yields. These findings contrast with those of wheat, soybean, and potato, which have shown slightly depressed yields at CO2 levels above 1200 μmol·mol–1.

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G.W. Stutte, C.L. Mackowiak, N.C. Yorio, R.M. Wheeler, and L.M. Ruffe

An experiment was conducted in the Biomass Production Chamber (BPC) at Kennedy Space Center to determine the feasibility of continuous steady-state production of potato (Solanum tuberosum L.). Plants were grown in a “batch” or continuous production mode using either 0.5 × modified Hoaglands or effluent from aerobically processed inedible potato biomass as a nutrient source. EC and pH were controlled to 0.12 S·m–1 and 5.8, respectively. The batch harvest occurred after 104 days and continuous harvest occurred every 26 days, with replanting occurring in the same solution. Continuous production on “aged” solution resulted in earlier tuber initiation, reduced plant height, and smaller canopies than the “batch” treatment. Planting density of the continuous treatment was increased from eight to 16 plants/m2. Because one quarter of the planting area was harvested and replanted every 26 days, a steady-state of canopy coverage between 60% to 75% of the chamber was maintained. Steady-state of CO2 fixation was also maintained in the continuous treatment. There was no effect on either quantum efficiency, tuber yield, or harvest index of the plants grown in continuous production. Although replanting into “aged” nutrient solution resulted in earlier tuber initiation and reduced plant size, the system reached a steady state of production, which is desirable for advanced life support system.

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C.L. Mackowiak, R.M. Wheeler, G.W. Stutte, N.C. Yorio, and L.M. Ruffe

Peanut (Arachis hypogaea L.) plants were grown hydroponically, using continuously recirculating nutrient solution. Two culture tray designs were tested; one tray design used only nutrient solution, while the other used a sphagnum-filled pod development compartment just beneath the cover and above the nutrient solution. Both trays were fitted with slotted covers to allow developing gynophores to reach the root zone. Peanut seed yields averaged 350 g·m-2 dry mass, regardless of tray design, suggesting that substrate is not required for hydroponic peanut production.

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N.C. Yorio, G.W. Stutte, D.S. DeVilliers, R.M. Wheeler, and R.L. Langhans

Bean (Phaseolus vulgaris L.) cv. Etna, a dry bean variety, and cv. Hystyle, a snap bean variety, were grown at 400 and 1200 μmol·m-2·s-1 CO2 to determine the effects of CO2 enrichment on plant growth and stomatal conductance. Plants were grown in controlled environment chambers for 70 days at each CO2 level using nutrient film technique hydroponics. An 18-h light/6-h dark photoperiod was maintained for each test, with a corresponding thermoperiod of 28 °C/24 °C and constant 65% RH. Diurnal stomatal conductance measurements were made with a steady-state porometer at 28 days after planting (DAP) and 49 DAP. As expected, plant growth and yield was consistently increased for each cultivar when plants were grown at 1200 μmol·m-2·s-1 CO2 compared to 400 μmol·m-2·s-1 CO2. Stomatal conductance measured during the light period showed an expected decrease for each cultivar when grown at 1200 μmol·m-2·s-1 CO2 compared to 400 μmol·m-2·s-1 CO2. However, during the dark period, stomatal conductance was higher for each cultivar grown at 1200 μmol·m-2·s-1 CO2. These results suggest a stomatal opening effect in the dark when plants are exposed to enriched levels of CO2. Tests are underway to investigate the effects of CO2 levels greater than 1200 μmol·m-2·s-1 on the growth and stomatal conductance of bean.

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N.C. Yorio, G.W. Stutte, G.D. Goins, D.S. de Villiers, and R.M. Wheeler

The effects of planting density and short-term changes in photoperiod on the growth and photosynthesis of bean (Phaseolus vulgaris L.) was investigated. Two cultivars of bean (cv. Etna, a dry bean variety; cv. Hystyle, a snap bean variety) were grown using nutrient film technique hydroponics in a walk-in growth chamber with a 12 h/12 h (light/dark) photoperiod and a corresponding thermoperiod of 28/24 °C (light/dark) and constant 65% relative humidity. Lighting for the chamber consisted of VHO fluorescent lamps and irradiance at canopy level was 400 μmol·m-2·s-1 PPF. For each cultivar, plants were grown at densities of 16 or 32 plants/m2. Short-term photoperiod changes were imposed during vegetative growth (21-29 DAP) and pod-fill (42-57 DAP). From the base 12 h/12h (light/dark) photoperiod, lighting in the chamber was cycled to provide 18 h/06 h (light/dark) or 24 h/0 h(continuous light) for 48 h. Diurnal single leaf net photosynthetic rates (Pn) and net assimilation vs. internal CO2 (Aci) measurements were taken during the short-term photoperiod adjustments. Results showed that there was no difference between cultivars or planting density with regard to total biomass or single leaf photosynthetic rates, but cv. Etna produced 35% more edible biomass than cv. Hystyle. Additionally, there was no effect of short-term photoperiod adjustment on single leaf Pn or Aci.

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R.M. Wheeler, C.L. Mackowiak, J.C. Sager, N.C. Yorio, W.M. Knott, and W.L. Berry

Two studies were conducted in which `Waldmann's Green' lettuce (Lactuca sativa L.) was grown hydroponically from seed to harvest in a large (20-m2), atmospherically closed growth chamber for the National Aeronautics and Space Administration's controlled ecological life support system (CELSS) program. The first study used metal-halide (MH) lamps [280 μmol·m-2·s-1 photosynthetic photon flux (PPF)], whereas the second used high-pressure sodium (HPS) lamps (293 μmol·m-2·s-1). Both studies used a 16-hour photoperiod, a constant air temperature (22 to 23C), and 1000 μmol·mol-1 CO2 during the light period. In each study, canopy photosynthesis and evapotranspiration (ET) rates were highly correlated to canopy cover, with absolute rates peaking at harvest (28 days after planting) at 17 μmol CO2/m2 per sec and 4 liters·m-2·day-1, respectively. When normalized for actual canopy cover, photosynthesis and ET rates per unit canopy area decreased with age (between 15 and 28 days after planting). Canopy cover increased earlier during the study with HPS lamps, and final shoot yields averaged 183 g fresh mass (FM)/plant and 8.8 g dry mass (DM)/plant. Shoot yields in the first study with MH lamps averaged 129 g FM/plant and 6.8 g DM/plant. Analysis of leaf tissue showed that ash levels from both studies averaged 22% and K levels ranged from 15% to 17% of tissue DM. Results suggest that lettuce should be easily adaptable to a CELSS with moderate lighting and that plant spacing or transplant schemes are needed to maximize canopy light interception and sustain efficient CO2 removal and water production.