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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↗

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↗

Environmental modification of yield and nutrient composition of 'Waldmann's Green' leaf lettuce

Leaf number, dry weight, and nutrient composition of Lactuca sativa L. cv. Waldmann's Green leaves were compared following 9 days of treatment in a controlled environment room under various combinations of photosynthetic photon flux (PPF:350 vs 800 micromoles m-2 s-1), atmospheric CO2 level (ambient vs 1500 micromoles mol-1), and single-strength (1X:15 mM) vs double-strength (2X:30 mM) nitrogen (N) as NO3- alone or as NH4(+) + NO3- (1:5 molar ratio). CO2 enrichment greatly enhanced leaf number under all PPF and N conditions, but increased leaf dry weight only at high PPF. Conditions favoring high photosynthesis enhanced leaf starch content 3-fold, and protein content increased as much as 64% with 2X NH4(+)+NO3-. Free sugar content was 6 to 9% of leaf dry weight for all treatment combinations, while fat was 1.5 to 3.5%. Ash content varied from 15 to 20% of leaf dry weight. Modified controlled environments can be used to enhance the nutritional content as well as the yield of crops to be used for life support in space-deployed, self-sustaining human habitats. Leaf lettuce is a useful model crop for demonstrating the potential of nutritional value added by environmental manipulation.

Non-NASA Center↗

Comparative induction of nitrate reductase by nitrate and nitrite in barley leaves

The comparative induction of nitrate reductase (NR) by ambient NO3- and NO2- as a function of influx, reduction (as NR was induced) and accumulation in detached leaves of 8-day-old barley (Hordeum vulgare L.) seedlings was determined. The dynamic interaction of NO3- influx, reduction and accumulation on NR induction was shown. The activity of NR, as it was induced, influenced its further induction by affecting the internal concentration of NO3-. As the ambient concentration of NO3- increased, the relative influences imposed by influx and reduction on NO3- accumulation changed with influx becoming a more predominant regulant. Significant levels of NO3- accumulated in NO2(-)-fed leaves. When the leaves were supplied cycloheximide or tungstate along with NO2-, about 60% more NO3- accumulated in the leaves than in the absence of the inhibitors. In NO3(-)-supplied leaves NR induction was observed at an ambient concentration of as low as 0.02 mM. No NR induction occurred in leaves supplied with NO2- until the ambient NO2- concentration was 0.5 mM. In fact, NR induction from NO2- solutions was not seen until NO3- was detected in the leaves. The amount of NO3- accumulating in NO2(-)-fed leaves induced similar levels of NR as did equivalent amounts of NO3- accumulating from NO3(-)-fed leaves. In all cases the internal concentration of NO3-, but not NO2-, was highly correlated with the amount of NR induced. The evidence indicated that NO3- was a more likely inducer of NR than was NO2-.

NASA Program CELSS↗

Expression of characteristics of ammonium nutrition as affected by pH of the root medium

To study the effect of root-zone pH on characteristic responses of NH4+ -fed plants, soybeans (Glycine max inverted question markL. inverted question mark Merr. cv. Ransom) were grown in flowing solution culture for 21 d on four sources of N (1.0 mol m-3 NO3-, 0.67 mol m-3 NO3- plus 0.33 mol m-3 NH4+, 0.33 mol m-3 NO3- plus 0.67 mol m-3 NH4+, and 1.0 mol m-3 NH4+) with nutrient solutions maintained at pH 6.0, 5.5, 5.0, and 4.5. Amino acid concentration increased in plants grown with NH4+ as the sole source of N at all pH levels. Total amino acid concentration in the roots of NH4+ -fed plants was 8 to 10 times higher than in NO3(-)-fed plants, with asparagine accounting for more than 70% of the total in the roots of these plants. The concentration of soluble carbohydrates in the leaves of NH4+ -fed plants was greater than that of NO3(-)-fed plants, but was lower in roots of NH4+ -fed plants, regardless of pH. Starch concentration was only slightly affected by N source or root-zone pH. At all levels of pH tested, organic acid concentration in leaves was much lower when NH4+ was the sole N source than when all or part of the N was supplied as NO3-. Plants grown with mixed NO3- plus NH4+ N sources were generally intermediate between NO3(-)- and NH4+ -fed plants. Thus, changes in tissue composition characteristic of NH4+ nutrition when root-zone pH was maintained at 4.5 and growth was reduced, still occurred when pH was maintained at 5.0 or above, where growth was not affected. The changes were slightly greater at pH 4.5 than at higher pH levels.

NASA Discipline Life Support Systems↗

Stimulation of nodulation in field peas (Pisum sativum) by low concentrations of ammonium in hydroponic culture

Although the inhibitory effects of high concentrations of mineral N (> 1.0 mM) on nodule development and function have often been studied, the effects of low, static concentrations of NH4+ (< 1.0 mM) on nodulation are unknown. In the present experiments we examine the effects of static concentrations of NH4+ at 0, 0.1 and 0.5 mM in flowing, hydroponic culture on nodule establishment and nitrogenase activity in field peas [Pisum sativum L. cv. Express (Svalof AB)] for the initial 28 days after planting (DAP). Peas grown in the presence of low concentrations of NH4+ had significantly greater nodule numbers (up to 4-fold) than plants grown without NH4+. Nodule dry weight per plant was significantly higher at 14, 21 and 28 DAP in plants grown in the presence of NH4+, but individual nodule mass was lower than in plants grown without NH4+. The nodulation pattern of the plants supplied with NH4+ was similar to that often reported for supernodulating mutants, however the plants did not express other growth habits associated with supernodulation. Estimates of N2 fixation indicate that the plus-NH4+ peas fixed as much or more N2 than the plants supplied with minus-NH4+ nutrient solution. There were no significant differences in nodule numbers, nodule mass or NH4+ uptake between the plants grown at the two concentrations of NH4+. Nodulation appeared to autoregulate by 14 DAP in the minus-NH4+ treatment. Plant growth and N accumulation in the minus-NH4+ plants lagged behind those of the plus-NH4+ treatments prior to N2 fixation becoming well established in the final week of the experiment. The plus-NH4+ treatments appeared not to elicit autoregulation and plants continued to initiate nodules throughout the experiment.

NASA Program CELSS↗