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Dry matter and nitrogen accumulation are not affected by superoptimal concentration of ammonium in flowing solution culture with pH control

While it is known that superoptimal concentrations of the nitrate (NO3-) ion in solution culture do not increase NO3- uptake or dry matter accumulation, the same is not known for the ammonium (NH4+) ion. An experiment was conducted utilizing flowing solution culture with pH control to investigate the influence of superoptimal NH4+ concentrations on dry matter, nitrogen (N), potassium (K), calcium (Ca), and magnesium (Mg) accumulation by nonnodulated soybean plants. Increasing the NH4+ concentration in solution from 1 to 10 mM did not affect dry matter or N accumulation. Accumulations of K, Ca, and Mg were slightly decreased with increased NH4+ concentration. The NH4+ uptake system, which is saturated at less than 1mM NH4+, is able to regulate uptake of NH4+ at concentrations as high as 10 mM.

NASA Discipline Life Support Systems↗

Effects of root-zone acidity on utilization of nitrate and ammonium in tobacco plants

Tobacco (Nicotiana tabacum L., cv. 'Coker 319') plants were grown for 28 days in flowing nutrient culture containing either 1.0 mM NO3- or 1.0 mM NH4+ as the nitrogen source in a complete nutrient solution. Acidities of the solutions were controlled at pH 6.0 or 4.0 for each nitrogen source. Plants were sampled at intervals of 6 to 8 days for determination of dry matter and nitrogen accumulation. Specific rates of NO3- or NH4+ uptake (rate of uptake per unit root mass) were calculated from these data. Net photosynthetic rates per unit leaf area were measured on attached leaves by infrared gas analysis. When NO3- [correction of NO-] was the sole nitrogen source, root growth and nitrogen uptake rate were unaffected by pH of the solution, and photosynthetic activity of leaves and accumulation of dry matter and nitrogen in the whole plant were similar. When NH4+ was the nitrogen source, photosynthetic rate of leaves and accumulation of dry matter and nitrogen in the whole plant were not statistically different from NO3(-) -fed plants when acidity of the solution was controlled at pH 6.0. When acidity for NH4(+) -fed plants was increased to pH 4.0, however, specific rate of NH4+ uptake decreased by about 50% within the first 6 days of treatment. The effect of acidity on root function was associated with a decreased rate of accumulation of nitrogen in shoots that was accompanied by a rapid cessation of leaf development between days 6 and 13. The decline in leaf growth rate of NH4(+) -fed plants at pH 4.0 was followed by reductions in photosynthetic rate per unit leaf area. These responses of NH4(+) -fed plants to increased root-zone acidity are characteristic of the sequence of responses that occur during onset of nitrogen stress.

NASA Discipline Life Support Systems↗

Potassium uptake supporting plant growth in the absence of AKT1 channel activity: Inhibition by ammonium and stimulation by sodium

A transferred-DNA insertion mutant of Arabidopsis that lacks AKT1 inward-rectifying K+ channel activity in root cells was obtained previously by a reverse-genetic strategy, enabling a dissection of the K+-uptake apparatus of the root into AKT1 and non-AKT1 components. Membrane potential measurements in root cells demonstrated that the AKT1 component of the wild-type K+ permeability was between 55 and 63% when external [K+] was between 10 and 1,000 microM, and NH4+ was absent. NH4+ specifically inhibited the non-AKT1 component, apparently by competing for K+ binding sites on the transporter(s). This inhibition by NH4+ had significant consequences for akt1 plants: K+ permeability, 86Rb+ fluxes into roots, seed germination, and seedling growth rate of the mutant were each similarly inhibited by NH4+. Wild-type plants were much more resistant to NH4+. Thus, AKT1 channels conduct the K+ influx necessary for the growth of Arabidopsis embryos and seedlings in conditions that block the non-AKT1 mechanism. In contrast to the effects of NH4+, Na+ and H+ significantly stimulated the non-AKT1 portion of the K+ permeability. Stimulation of akt1 growth rate by Na+, a predicted consequence of the previous result, was observed when external [K+] was 10 microM. Collectively, these results indicate that the AKT1 channel is an important component of the K+ uptake apparatus supporting growth, even in the "high-affinity" range of K+ concentrations. In the absence of AKT1 channel activity, an NH4+-sensitive, Na+/H+-stimulated mechanism can suffice.

Non-NASA Center↗

Solubility and Cation Exchange Properties of Synthetic Hydroxyapatite and Clinoptilolite Mixtures

A zeoponic plant growth system is defined as the cultivation of plants in artificial soils, which have zeolites as a major component. These systems: 1) can serve as a controllable and renewable fertilization system to provide plant growth nutrients; 2) can mitigate the adverse effects of contamination due to leaching of highly soluble and concentrated fertilizers; and 3) are being considered as substrates for plant growth in regenerative life-support systems for long-duration space missions. Batch-equilibrium studies of the dissolution and ion-exchange properties of mixtures of naturally-occurring Wyoming clinoptilolite (a zeolite) exchanged with K(+) or NH4(+); and synthetic hydroxyapatite were conducted to determine: 1) the plant availability of the macro-nutrients NH4-N, P, K, Ca, and Mg and 2) the effects of varying the clinoptilolite to hydroxyapatite ratio and the ratio of exchangeable cations (K(+) vs. NH4(+)) on clinoptilolite extraframework sites. The nutrients NH4-N (19.7 to 73.6 mg L(sup -1), P (0.57 to 14.99 mg L(sup- 1), K (14.8 to 104.9 mg L(sup -1), and Mg (0.11 to 6.68mg L(sup -1) are available to plants at sufficient levels. Solution Ca concentrations (0.47 to 3.40 mg L(sup -1) are less than optimal. Solution concentrations of NH4(+), K(+), Ca(2+), and Mg(2+) all decreased with increasing clinoptilolite to hydroxyapatite ratio in the sample. Solution concentrations of phosphorous initially increased, reached a maximum value and then decreased with increasing clinoptilolite to hydroxyapatite ratio in the sample. The NH4(+) -exchanged clinoptilolite is more efficient in dissolving synthetic hydroxyapatite than the K(+) -exchanged clinoptilolite. This suggests that NH4(+), which is less selective at clinoptilolite extraframework sites than K(+) is exchanged more readily by Ca(2+) and thereby enhances the dissolution of the synthetic hydroxyapatite.

Beiersdorfer, Raymond E.↗

Response of potatoes to nitrogen concentrations differ with nitrogen forms

Two separate experiments were conducted to investigate plant growth and mineral composition of potatoes (Solanum tuberosum L.) at varied solution concentrations of nitrate (NO3-) and ammonium (NH4+). Each experiment evaluated five nitrogen (N) concentrations of 0.5, 2, 4, 8, and 12 mM, which were maintained with a non-recirculating nutrient film system in controlled environment. Plants were harvested on day 42 with NO3-; and day 35 with NH4+ after transplanting of tissue culture plantlets, and growth measurements were taken as leaf area, tuber number, and dry weights of different parts. With NO3-, plant growth was greatest and similar at 2, 4, and 8 mM of N whereas with NH4+, plant growth was best only at 2 and 4 mM of N. At 12 mM of N, plants exhibited interveinal ammonium toxicity with NH4+ nutrition, but healthy growth appearance with NO3- nutrition. With either N form, total N concentrations in tissues tended to increase with increased N concentrations, and tissue phosphorus (P) concentrations were reduced at 0.5 and 2 mM of N. Tissue concentrations of calcium (Ca), magnesium (Mg), and sulfur (S) changed only slightly at particular N concentrations, yet changed substantially with different N forms. The data indicate that the optimal ranges of N concentrations in both solution and tissues are wider and higher with NO3- than with NH4+ nutrition, and thus a careful control of NH4+ concentrations is necessary to minimize possible ammonium toxicity to potato plants.

NASA Discipline Life Support Systems↗

Solubility and cation exchange in phosphate rock and saturated clinoptilolite mixtures

Mixtures of zeolite and phosphate rock (PR) have the potential to provide slow-release fertilization of plants in synthetic soils by dissolution and ion-exchange reactions. This study was conducted to examine solubility and cation-exchange relationships in mixtures of PR and NH4- and K-saturated clinoptilolite (Cp). Batch-equilibration experiments were designed to investigate the effect of PR source, the proportion of exchangeable K and NH4, and the Cp to PR ratio on solution N, P, K, and Ca concentrations. The dissolution and cation-exchange reactions that occurred after mixing NH4- and K-saturated Cp with PR increased the solubility of the PR and simultaneously released NH4 and K into solution. The more reactive North Carolina (NC) PR rendered higher solution concentrations of NH4 and K when mixed with Cp than did Tennessee (TN) PR. Solution P concentrations for the Cp-NC PR mixture and the Cp-TN PR mixture were similar. Solution concentrations of N, P, K, and Ca and the ratios of these nutrients in solution varied predictably with the type of PR, the Cp/PR ratio, and the proportions of exchangeable K and NH4 on the Cp. Our research indicated that slow-release fertilization using Cp/PR media may provide adequate levels of N, P, and K to support plant growth. Solution Ca concentrations were lower than optimum for plant growth.

NASA Discipline Number 61-10↗

Utilization of ammonium as a nitrogen source: effects of ambient acidity on growth and nitrogen accumulation by soybean

Dry matter accumulation of plants utilizing NH4+ as the sole nitrogen source generally is less than that of plants receiving NO3- unless acidity of the root-zone is controlled at a pH of about 6.0. To test the hypothesis that the reduction in growth is a consequence of nitrogen stress within the plant in response to effects of increased acidity during uptake of NH4+ by roots, nonnodulated soybean plants (Glycine max [L.] Merr. cv Ransom) were grown for 24 days in flowing nutrient culture containing 1.0 millimolar NH4+ as the nitrogen source. Acidities of the culture solutions were controlled at pH 6.1, 5.1, and 4.1 +/- 0.1 by automatic additions of 0.01 N H2SO4 or Ca(OH)2. Plants were sampled at intervals of 3 to 4 days for determination of dry matter and nitrogen accumulation. Rates of NH4+ uptake per gram root dry weight were calculated from these data. Net CO2 exchange rates per unit leaf area were measured on attached leaves by infrared gas analysis. When acidity of the culture solution was increased from pH 6.1 to 5.1, dry matter and nitrogen accumulation were reduced by about 40% within 14 days. Net CO2 exchange rates per unit leaf area, however, were not affected, and the decreased growth was associated with a reduction in rates of appearance and expansion of new leaves. The uptake rates of NH4+ per gram root were about 25% lower throughout the 24 days at pH 5.1 than at 6.1. A further increase in solution acidity from pH 5.1 to 4.1 resulted in cessation of net dry matter production and appearance of new leaves within 10 days. Net CO2 exchange rates per unit leaf area declined rapidly until all viable leaves had abscised by 18 days. Uptake rates of NH4+, which were initially about 50% lower at pH 4.1 than at 6.1 continued to decline with time of exposure until net uptake ceased at 10 days. Since these responses also are characteristic of the sequence of responses that occur during onset and progression of a nitrogen stress, they corroborate our hypothesis.

NASA Discipline Number 61-10↗

Effect of ammonium sulfate, ammonium chloride and root-zone acidity on inorganic ion content of tobacco

Tobacco plants (Nicotiana tabacum L. cv NC82) were supplied with (NH4)2SO4 or NH4Cl at root-zone pH of 6.0 and 4.5 in hydroponic culture for 28 days. Dry matter accumulation, total N and C content, and leaf area and number were not affected by the NH4+ source or root-zone pH. Plants supplied with NH4Cl accumulated up to 1.2 mM Cl g DW-1, but accumulated 37% less inorganic H2PO4- and 47% less SO4(2-) than plants supplied with (NH4)2SO4. The large Cl- accumulation resulted in NH4Cl- supplied plants having a 31% higher inorganic anion (NO3-, H2, PO4-, SO4(2-), and Cl-) charge. This higher inorganic anion charge in the NH4Cl-supplied plants was balanced by a similar increase in K+ charge. Plants supplied with NH4Cl accumulated greater concentrations of Cl- in leaves (up to 5.1% of DW) than plants supplied with (NH4)2SO4 (less than -% DW). Despite the high Cl- concentration of leaves in NH4Cl supplied plants, these plants showed no symptoms of Cl- toxicity. This demonstrates that toxicity symptoms are not due solely to an interaction between high Cl- concentration in tissue and NH4+ nutrition. The increase in root-zone acidity to pH 4.5 from 6.0 did not induce toxicity symptoms.

NASA Program CELSS↗

A simulation experiment of photochemical reactions in the mesosphere

An ionospheric simulation experiment has been performed in a large vacuum chamber. The chamber is filled with NO and other gases including N2, O2, CO2, NH3 and H2O in the pressure range of 0.01 torr. A lamp which produces photons at 1236- and 1165-A by means of microwave discharge in krypton is utilized as an ionization source. In addition to 30+ large quantities of the water cluster ions 55+, H3O(+).(H2O)2, 73+, H3O(+).(H2O)3 and 91+, H3O(+).(H2O)4 were observed when nitric oxide and water were present. This closely approximates the condition of the terrestrial D region. After long periods of UV irradiation 74+ and 104+ ions grow in intensity. These ions are tentatively identified as NO(+).N2O and NO(+).NO.N2O. In addition the series 18+, 36+, 54+, and 72+ is detected which can be labeled NH4(+), NH4(+).(H2O), NH4(+).(H2O)2 and NH4(+).(H2O)3. These same species of ions are observed with the introduction of ammonia into the chamber. Presumably both N2O and NH3 are products of the photolysis.

Fugono, N.↗

Quantitative reflectance spectroscopy of buddingtonite from the Cuprite mining district, Nevada

Buddingtonite, an ammonium-bearing feldspar diagnostic of volcanic-hosted alteration, can be identified and, in some cases, quantitatively measured using short-wave infrared (SWIR) reflectance spectroscopy. In this study over 200 samples from Cuprite, Nevada, were evaluated by X ray diffraction, chemical analysis, scanning electron microscopy, and SWIR reflectance spectroscopy with the objective of developing a quantitative remote-sensing technique for rapid determination of the amount of ammonium or buddingtonite present, and its distribution across the site. Based upon the Hapke theory of radiative transfer from particulate surfaces, spectra from quantitative, physical mixtures were compared with computed mixture spectra. We hypothesized that the concentration of ammonium in each sample is related to the size and shape of the ammonium absorption bands and tested this hypothesis for samples of relatively pure buddingtonite. We found that the band depth of the 2.12-micron NH4 feature is linearly related to the NH4 concentration for the Cuprite buddingtonite, and that the relationship is approximately exponential for a larger range of NH4 concentrations. Associated minerals such as smectite and jarosite suppress the depth of the 2.12-micron NH4 absorption band. Quantitative reflectance spectroscopy is possible when the effects of these associated minerals are also considered.

Felzer, Benjamin↗

Evaluation of an Anaerobic Digestion System for Processing CELSS Crop Residues for Resource Recovery

Three bioreactors, connected in series, were used to process CELSS potato residues for recovery of resources. The first stage was an anaerobic digestor (8 L working volume; cow rumen contents inoculum; fed-batch; 8 day retention time; feed rate 25 gdw/day) that converted 33% of feed (dry weight loss) to CO2 and "volatile fatty acids" (vfa, 83:8:8 mmolar ratio acetic:propionic:butyric). High nitrate-N in the potato residue feed was absent in the anaerobic effluent, with a high portion converted to NH4(+)-N and the remainder unaccounted and probably lost to denitrification and NH4(+) volatilization. Liquid anaerobic effluent was fed to an aerobic, yeast biomass production vessel (2 L volume; Candida ingens inoculum; batch [pellicle] growth; 2 day retention time) where the VFAs and some NH4(+)-N were converted into yeast biomass. Yeast yields accounted for up to 8% of potato residue fed into the anaerobic bioreactor. The third bioreactor (0.5 L liquid working volume; commercial nitrifier inoculum; packed-bed biofilm; continuous yeast effluent feed; recirculating; constant volume; 2 day hydraulic retention time) was used to convert successfully the remaining NH4(+)-N into nitrate-N (preferred form of N for CELSS crop production) and to remove the remaining degradable soluble organic carbon. Effluents from the last two stages were used for partial replenishment of minerals for hydroponic potato production.

Strayer, R. F.↗

Nitrification in a zeoponic substrate

Clinoptilolite is a zeolite mineral with high cation exchange capacity used in zeoponic substrates that have been proposed as a solid medium for growing plants or as a fertilizer material. The kinetics of nitrification has not been measured for NH4+ saturated zeoponic substrate. Experiments were conducted to evaluate the production of NO2- and NO3-, and nitrifier populations in zeoponic substrates. Small columns were filled with zeoponic substrate inoculated with a commercial inoculum or soil enrichment culture of nitrifying bacteria. In addition to column studies, a growth chamber study was conducted to evaluate the kinetics of nitrification in zeoponic substrates used to grow radishes (Raphanus sativus L.). The zeoponic substrate provided a readily available source of NH4+, and nitrifying bacteria were active in the substrate. Ammonium oxidation rates in column studies ranged from 5 to 10 micrograms N g-1 substrate h-1, and NO2- oxidation rates were 2 to 9.5 micrograms N g-1 substrate h-1. Rates determined from the growth chamber study were approximately 1.2 micrograms N g-1 substrate h-1. Quantities of NH4+ oxidized to NO2- and NO3- in inoculated zeoponic substrate were in excess of plant up-take. Acidification as a result of NH4+ oxidation resulted in a pH decline, and the zeoponic substrate showed limited buffering capacity.

NASA Discipline Life Support Systems↗

Plant Productivity and Characterization of Zeoponic Substrates after Three Successive Crops of Radish (Raphanus sativus L.)

The National Aeronautics and Space Administration (NASA) has developed a zeolite-based synthetic substrate, termed zeoponics. The zeoponic substrate (consisting of NH4(-) and K-exchanged clinoptilolite, synthetic apatite, and dolomite) provides all of the plant-essential nutrients through mineral dissolution and ion exchange, with only the addition of water. Previous studies have shown high productivity of wheat in zeoponic substrates; however, no experiments have been conducted on other crops. The objective of this study was to determine the productivity and nutrient uptake of radish (Raphanus sativus L.) grown in zeoponic substrates with three successive crops in the same substrate. Radish was chosen because of its sensitivities to NH4(+). Average fresh weights of edible roots were similar for radish grown in zeoponic substrates watered with deionized H2O (10.97 g/plant) and in potting mix control substrate irrigated with nutrient solution (10.92 g/plant). Average fresh weight production of edible roots for radish grown in same zeoponic substrate increased in yield over time with the lowest yield in the first crop (7.10 g/plant) and highest in the third crop (13.90 g/plant). The Ca plant tissue levels in radishes (1.8-2.9 wt. %) grown in zeoponic substrates are lower than the suggested sufficient range of 3.0-4.5 wt. % Ca; however, the Ca level is highest (2.9 wt. %) in radishes grown in the third crop in the same zeoponic substrates. The higher radish yield in the third crop was attributed to a reduction in an NH4(-) induced Ca deficiency that has been previously described for wheat grown in zeoponic substrates. The P levels in plant tissues of radish grown in the zeoponic substrates ranged from 0.94-1.15 wt. %; which is slightly higher than the sufficient levels of 0.3-0.7 wt. %. With the exception of Ca and P, other macronutrient and micronutrient levels in radish grown in zeoponic substrates were well within the recommended sufficient ranges. After three successive crops of radish growth, the zeoponic substrates had 52% of the original NH4(-)N and 78% of the original K remaining on zeolite exchange sites. Zeoponic substrates are capable of long-term productivity of radishes for space.

Gruener, J. E.↗

Plant Growth Experiments in Zeoponic Substrates: Applications for Advanced Life Support Systems

A zeoponic plant-growth system is defined as the cultivation of plants in artificial soils, which have zeolites as a major component (Allen and Ming, 1995). Zeolites are crystalline, hydrated aluminosilicate minerals that have the ability to exchange constituent cations without major change of the mineral structure. Recently, zeoponic systems developed at the National Aeronautics and Space Administration (NASA) slowly release some (Allen et at., 1995) or all of the essential plant-growth nutrients (Ming et at., 1995). These systems have NH4- and K-exchanged clinoptilolite (a natural zeolite) and either natural or synthetic apatite (a calcium phosphate mineral). For the natural apatite system, Ca and P were made available to the plant by the dissolution of apatite. Potassium and NH4-N were made available by ion-exchange reactions involving Ca(2+) from apatite dissolution and K(+) and NH4(+) on zeolitic exchange sites. In addition to NH4-N, K, Ca, and P, the synthetic apatite system also supplied Mg, S, and other micronutrients during dissolution (Figure 1). The overall objective of this research task is to develop zeoponic substrates wherein all plant growth nutrients are supplied by the plant growth medium for several growth seasons with only the addition of water. The substrate is being developed for plant growth in Advanced Life Support (ALS) testbeds (i.e., BioPLEX) and microgravity plant growth experiments. Zeoponic substrates have been used for plant growth experiments on two Space Shuttle flight experiments (STS-60; STS-63; Morrow et aI., 1995). These substrates may be ideally suited for plant growth experiments on the International Space Station and applications in ALS testbeds. However, there are several issues that need to be resolved before zeoponics will be the choice substrate for plant growth experiments in space. The objective of this paper is to provide an overview on recent research directed toward the refinement of zeoponic plant growth substrates.

Ming, Douglas W.↗

A kinetic study of the interaction between atomic oxygen and aerosols

This study was concerned with the effects of NH4Cl and (NH4)2SO4 aerosols on the kinetics of disappearance of atomic oxygen. Atomic oxygen was generated by a 2.45-GHz microwave discharge and the kinetics of disappearance measured in a fast flow system using NO2 titration. Values of the recombination coefficient for heterogeneous wall recombination were determined for clean, H2SO4-coated, and (NH4)2SO4-coated Pyrex to be 0.000050, 0.000020, and 0.000019, respectively. A rapid exothermic chemical reaction was found to occur between atomic oxygen and an NH4Cl wall coating; the products were NH3, NO, H2O, and HCl. The NH4Cl aerosol was generated by gas phase reaction of NH3 with HCl. The aerosol particles were approximately spherical and nearly monodisperse with a mean diameter of 1.6 plus or minus 0.2 micron. The rate constant for the disappearance of atomic oxygen in the presence of NH4Cl aerosol was measured. No significant decrease was observed in the rate of disappearance of atomic oxygen in the presence of an (NH4)2SO4 aerosol at a concentration of 285 mg per cu m.

Akers, F. I.↗

Metastable Solution Thermodynamic Properties and Crystal Growth Kinetics

The crystal growth rates of NH4H2PO4, KH2PO4, (NH4)2SO4, KAl(SO4)2 central dot 12H2O, NaCl, and glycine and the nucleation rates of KBr, KCl, NaBr central dot 2H2O, (NH4)2Cl, and (NH4)2SO4 were expressed in terms of the fundamental driving force of crystallization calculated from the activity of supersaturated solutions. The kinetic parameters were compared with those from the commonly used kinetic expression based on the concentration difference. From the viewpoint of thermodynamics, rate expressions based on the chemical potential difference provide accurate kinetic representation over a broad range of supersaturation. The rates estimated using the expression based on the concentration difference coincide with the true rates of crystallization only in the concentration range of low supersaturation and deviate from the true kinetics as the supersaturation increases.

Kim, Soojin↗

Nitrogen recycling during phenylpropanoid metabolism in sweet potato tubers

In the first step of the phenylpropanoid metabolic pathway, L-phenylalanine (L-Phe) is deaminated to form E-cinnamate, in a conversion catalyzed by phenylalanine ammonia-lyase (PAL; EC 4.3.1.5). The metabolic fate of the ammonium ion (NH4+) produced in this reaction was investigated in sweet potato (Ipomoea batatas) tuber discs. [15N]-Labeled substrates including L-Phe, in the presence or absence of specific enzyme inhibitors, were administered to sweet potato discs in light under aseptic conditions. 15N-Nuclear magnetic resonance spectroscopic analyses revealed that the 15NH4+ liberated during the PAL reaction is first incorporated into the amide nitrogen of L-glutamine (L-Gln) and then into L-glutamate (L-Glu). These results extend our previous observations in pine and potato that PAL-generated NH4+ is assimilated by the glutamine synthetase (GS; EC 6.3.1.2)/glutamate synthase (GOGAT; EC 1.4.1.13) pathway, with the NH4+ so formed ultimately being recycled back to L-Phe via L-Glu as aminoreceptor and donor.

Non-NASA Center↗

Somatic embryogenesis of carrot in hormone-free medium: external pH control over morphogenesis

Cultures of preglobular stage proembryos (PGSPs) were initiated from mechanically wounded mature zygotic embryos of carrot, Daucus carota, on a hormone-free, semisolid medium. These PGSPs have been maintained and multiplied for extended periods without their progression into later embryo stages on the same hormone-free medium containing 1 mM NH4+ as the sole nitrogen source. Sustained maintenance of cultures comprised exclusively of PGSPs was dependent on medium pH throughout the culture period. Best growth and multiplication of PGSP cultures occurred when the pH of unbuffered, hormone-free medium fell from 4.5 to 4 over a 2-week period or when buffered medium was titrated to pH 4. If the hormone-free medium was buffered to sustain a pH at or above 4.5, PGSPs developed into later embryo stages. Maintenance with continuous multiplication of PGSPs occurred equally well on medium containing NH4+ or NH4+ and NO3-, but growth was poor with NO3- alone. Additional observations on the effects of medium components such as various nitrogen sources and levels, sucrose concentration, semisolid supports, type of buffer, borate concentration, activated charcoal, and initial pH that permit optimum maintenance of the PGSPs or foster their continued developmental progression into mature embryos and plantlets are reported. The influence of the pH of the hormone-free medium as a determinant in maintaining cultures as PGSPs or allowing their continued embryonic development are unequivocally demonstrated by gross morphology, scanning electron microscopy, and histological preparations.

NASA Discipline Number 29-20↗