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Expansion and photosynthetic rate of leaves of soybean plants during onset of and recovery from nitrogen stress

This study reports on the effects of nitrogen stress and restoration of nitrogen availability after a period of stress on expansion and photosynthetic rate of soybean leaves of differing maturity. We hypothesized that nitrogen resupply would lead to additional accumulation of reduced nitrogen in the leaves and, ultimately, resumption of leaf initiation and expansion and photosynthetic activity. In continuously nitrogen-stressed plants, expansion of middle trifoliolates of main-stem trifoliates and leaf area at full expansion were severely restricted. Leaves showing the greatest effects were initiated after removal of nitrogen. When the reduced nitrogen concentration in mature leaves of continuously stressed plants fell below 9 mg dm-2, the photosynthetic rate per unit leaf decreased rapidly, reaching a minimum of ca. 6-8 mg dm-2 h-1. The older mature leaves tended to abscise at this point, while the youngest leaves remained on the plant and continued to photosynthesize slowly. When nitrogen was resupplied, leaf expansion and final leaf area increased. Leaf initiation was also stimulated as reduced nitrogen levels rose in the leaves. Photosynthetic rates of the oldest and youngest pair of mature leaves returned to values comparable to those of similar-aged leaves of nonstressed soybean plants.

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↗

Do the higher oxidation states of the photosynthetic O2-evolving system contain bound H2O?

A modified mass spectrometer was used to determine whether the higher oxidation states of the photosynthetic O2-evolving system contain substrate water that is not freely exchangeable with the external medium. Our data indicated that the higher oxidation states contain no appreciable bound, non-exchangeable H2O. This suggests that H2O oxidation takes place via a rapid, concerted, all-or-none mechanism rather than by a mechanism involving stable, partially oxidized, H2O-derived intermediates. These findings set definite constraints on possible mechanisms of O2 evolution.

NASA Discipline Life Support Systems↗

Characteristics of injury and recovery of net NO3- transport of barley seedlings from treatments of NaCl

The nature of the injury and recovery of nitrate uptake (net uptake) from NaCl stress in young barley (Hordeum vulgare L, var CM 72) seedlings was investigated. Nitrate uptake was inhibited rapidly by NaCl, within 1 minute after exposure to 200 millimolar NaCl. The duration of exposure to saline conditions determined the time of recovery of NO3- uptake from NaCl stress. Recovery was dependent on the presence of NO3- and was inhibited by cycloheximide, 6-methylpurine, and cerulenin, respective inhibitors of protein, RNA, and sterol/fatty acid synthesis. These inhibitors also prevented the induction of the NO3- uptake system in uninduced seedlings. Uninduced seedlings exhibited endogenous NO3- transport activity that appeared to be constitutive. This constitutive activity was also inhibited by NaCl. Recovery of constitutive NO3- uptake did not require the presence of NO3-.

Non-NASA Center↗

Assessment of an apparent relationship between availability of soluble carbohydrates and reduced nitrogen during floral initiation in tobacco

Daily relative accumulation rate of soluble carbohydrates (RARS) and reduced nitrogen (RARN) in the shoot, as estimates of source strength, were compared with daily relative growth rates (RGR) of the shoot, as an estimate of sink demand, during floral transformation in apical meristems of tobacco (Nicotiana tabacum 'NC 2326') grown at day/night temperatures of 18/14, 22/18, 26/22, 30/26, and 34/30 C. Source strength was assumed to exceed sink demand for either carbohydrates or nitrogen when the ratio of RARS/RGR or RARN/RGR was greater than unity, and sink demand was assumed to exceed source strength when the ratio was less than unity. Time of floral initiation, which was delayed up to 21 days with increases in temperature over the experimental range, was associated with intervals in which source strength of either carbohydrate or nitrogen exceeded sink demand, while sink demand for the other exceeded source strength. Floral initiation was not observed during intervals in which source strengths of both carbohydrates and nitrogen were greater than or less than sink demand. These results indicate that floral initiation is responsive to an imbalance in the relative availabilities of carbohydrate and nitrogen.

NASA Discipline Life Support Systems↗

Fourier transform-infrared spectroscopic methods for microbial ecology: analysis of bacteria, bacteria-polymer mixtures and biofilms

Fourier transform-infrared (FT-IR) spectroscopy has been used to rapidly and nondestructively analyze bacteria, bacteria-polymer mixtures, digester samples and microbial biofilms. Diffuse reflectance FT-IR (DRIFT) analysis of freeze-dried, powdered samples offered a means of obtaining structural information. The bacteria examined were divided into two groups. The first group was characterized by a dominant amide I band and the second group of organisms displayed an additional strong carbonyl stretch at approximately 1740 cm-1. The differences illustrated by the subtraction spectra obtained for microbes of the two groups suggest that FT-IR spectroscopy can be utilized to recognize differences in microbial community structure. Calculation of specific band ratios has enabled the composition of bacteria and extracellular or intracellular storage product polymer mixtures to be determined for bacteria-gum arabic (amide I/carbohydrate C-O approximately 1150 cm-1) and bacteria-poly-beta-hydroxybutyrate (amide I/carbonyl approximately 1740 cm-1). The key band ratios correlate with the compositions of the material and provide useful information for the application of FT-IR spectroscopy to environmental biofilm samples and for distinguishing bacteria grown under differing nutrient conditions. DRIFT spectra have been obtained for biofilms produced by Vibrio natriegens on stainless steel disks. Between 48 and 144 h, an increase in bands at approximately 1440 and 1090 cm-1 was seen in FT-IR spectra of the V. natriegens biofilm. DRIFT spectra of mixed culture effluents of anaerobic digesters show differences induced by shifts in input feedstocks. The use of flow-through attenuated total reflectance has permitted in situ real-time changes in biofilm formation to be monitored and provides a powerful tool for understanding the interactions within adherent microbial consortia.

NASA Discipline Number 61-10↗

Exploring the limits of crop productivity. I. Photosynthetic efficiency of wheat in high irradiance environments

The long-term vegetative and reproductive growth rates of a wheat crop (Triticum aestivum L.) were determined in three separate studies (24, 45, and 79 days) in response to a wide range of photosynthetic photon fluxes (PPF, 400-2080 micromoles per square meter per second; 22-150 moles per square meter per day; 16-20 hour photoperiod) in a near-optimum, controlled-environment. The CO2 concentration was elevated to 1200 micromoles per mole, and water and nutrients were supplied by liquid hydroponic culture. An unusually high plant density (2000 plants per square meter) was used to obtain high yields. Crop growth rate and grain yield reached 138 and 60 grams per square meter per day, respectively; both continued to increase up to the highest integrated daily PPF level, which was three times greater than a typical daily flux in the field. The conversion efficiency of photosynthesis (energy in biomass/energy in photosynthetic photons) was over 10% at low PPF but decreased to 7% as PPF increased. Harvest index increased from 41 to 44% as PPF increased. Yield components for primary, secondary, and tertiary culms were analyzed separately. Tillering produced up to 7000 heads per square meter at the highest PPF level. Primary and secondary culms were 10% more efficient (higher harvest index) than tertiary culms; hence cultural, environmental, or genetic changes that increase the percentage of primary and secondary culms might increase harvest index and thus grain yield. Wheat is physiologically and genetically capable of much higher productivity and photosynthetic efficiency than has been recorded in a field environment.

NASA Discipline Number 61-10↗

Inhibition of nitrate transport by anti-nitrate reductase IgG fragments and the identification of plasma membrane associated nitrate reductase in roots of barley seedlings

Membrane associated nitrate reductase (NR) was detected in plasma membrane (PM) fractions isolated by aqueous two-phase partitioning from barley (Hordeum vulgare L. var CM 72) roots. The PM associated NR was not removed by washing vesicles with 500 millimolar NaCl and 1 millimolar EDTA and represented up to 4% of the total root NR activity. PM associated NR was stimulated up to 20-fold by Triton X-100 whereas soluble NR was only increased 1.7-fold. The latency was a function of the solubilization of NR from the membrane. NR, solubilized from the PM fraction by Triton X-100 was inactivated by antiserum to Chlorella sorokiniana NR. Anti-NR immunoglobulin G fragments purified from the anti-NR serum inhibited NO3- uptake by more than 90% but had no effect on NO2- uptake. The inhibitory effect was only partially reversible; uptake recovered to 50% of the control after thorough rinsing of roots. Preimmune serum immunoglobulin G fragments inhibited NO3- uptake 36% but the effect was completely reversible by rinsing. Intact NR antiserum had no effect on NO3- uptake. The results present the possibility that NO3- uptake and NO3- reduction in the PM of barley roots may be related.

NASA Discipline Number 61-10↗

Temperature and photoperiod responses of soybean embryos cultured in vitro

Temperature and photoperiod each have direct effects on growth rate of excised embryos of soybean (Glycine max (L.) Merrill). To determine if the effects of photoperiod are altered by temperature, embryos of 'Ransom II' were cultured in vitro at 18, 24, and 30 degrees C under photoperiod durations of 12 and 18 h at an irradiance of 9 W m-2 (700 to 850 nm) and a photosynthetic photon flux density of 58 micromoles m-2 s-1 (400 to 700 nm). Accumulation rates of fresh and dry weight were greater under 18-h than 12-h photoperiods over the entire range of temperature. Water content of the culture embryos was not affected by photoperiod but was greater at 18 and 30 than 24 degrees C. The accumulation rate of dry weight increased from 18 to 26 but declined at 30 degrees C.

NASA Program CELSS↗

Characterization and subcellular localization of aminopeptidases in senescing barley leaves

Four aminopeptidases (APs) were separated using native polyacrylamide gel electrophoresis of cell-free extracts and the stromal fractions of isolated chloroplasts prepared from primary barley (Hordeum vulgare L., var Numar) leaves. Activities were identified using a series of aminoacyl-beta-naphthylamide derivatives as substrates. AP1, 2, and 3 were found in the stromal fraction of isolated chloroplasts with respective molecular masses of 66.7, 56.5, and 54.6 kilodaltons. AP4 was found only in the cytoplasmic fraction. No AP activity was found in vacuoles of these leaves. It was found that 50% of the L-Leu-beta-naphthylamide and 25% of the L-Arg-beta-naphthylamide activities were localized in the chloroplasts. Several AP activities were associated with the membranes of the thylakoid fraction of isolated chloroplasts. AP1, 2, and 4 reacted against a broad range of substrates, whereas AP3 hydrolyzed only L-Arg-beta-naphthylamide. Only AP2 hydrolyzed L-Val-beta-naphthylamide. Since AP2 and AP3 were the only ones reacting against Val-beta-naphthylamide and Arg-beta-naphthylamide, respectively, several protease inhibitors were tested against these substrates using a stromal fraction from isolated chloroplasts as the source of the two APs. Both APs were sensitive to both metallo and sulfhydryl type inhibitors. Although AP activity decreased as leaves senesced, no new APs appeared on gels during senescence and none disappeared.

NASA Discipline Number 61-10↗

Lipid content and fatty acid composition of green algae Scenedesmus obliquus grown in a constant cell density apparatus

The lipids of alga Scenedesmus obliquus grown under controlled conditions were separated and fractionated by column and thin-layer chromatography, and fatty acid composition of each lipid component was studied by gas-liquid chromatography (GLC). Total lipids were 11.17%, and neutral lipid, glycolipid and phospholipid fractions were 7.24%, 2.45% and 1.48% on a dry weight basis, respectively. The major neutral lipids were diglycerides, triglycerides, free sterols, hydrocarbons and sterol esters. The glycolipids were: monogalactosyl diglyceride, digalactosyl diglyceride, esterified sterol glycoside, and sterol glycoside. The phospholipids included: phosphatidyl choline, phosphatidyl glycerol and phosphatidyl ethanolamine. Fourteen fatty acids were identified in the four lipid fractions by GLC. The main fatty acids were C18:2, C16:0, C18:3(alpha), C18:1, C16:3, C16:1, and C16:4. Total unsaturated fatty acid and essential fatty acid compositions of the total algal lipids were 80% and 38%, respectively.

NASA Discipline Life Support Systems↗

Growth and tuberization of potato (Solanum tuberosum L.) under continuous light

The growth and tuberization of potatoes (Solanum tuberosum L.) maintained for 6 weeks under four different regimes of continuous irradiance were compared to plants given 12 hours light and 12 hours dark. Treatments included: (a) continuous photosynthetic photon flux of 200 micromoles per square meter per second cool-white fluorescent (CWF); (b) continuous 400 micromoles per square meter per second CWF; (c) 12 hours 400 micromoles per square meter per second CWF plus 12 hours dim CWF at 5 micromoles per square meter per second; (d) 12 hours [400] micromoles per square meter per second CWF plus 12 hours dim incandescent (INC) at 5 micromoles per square meter per second and a control treatment of 12 hours light at 400 micromoles per square meter per second CWF and 12 hours dark. The study included five cultivars ranging from early- to late-season types: 'Norland,' 'Superior,''Norchip,' 'Russet Burbank,' and 'Kennebec,' Tuber development progressed well under continuous irradiation at 400 micromoles per square meter per second and under 12 hours irradiance and 12 hours dark, while tuber development was suppressed in all other light treatments. Continuous irradiation at 200 or 400 micromoles per square meter per second resulted in severe stunting and leaf malformation on 'Superior' and 'Kennebec' plants, but little or no injury and vigorous shoot growth in the other cultivars. No injury or stunting were apparent under 12-dim light or 12-dark treatments. Plants given 12 hours dim INC showed significantly greater stem elongation but less total biomass than plants in other treatments. The continuous light encouraged shoot growth over tuber growth but this trend was overridden by providing a high irradiance level. The variation among cultivars for tolerance to continuous lighting indicates that potato may be a useful species for photoinhibition studies.

NASA Program CELSS↗

A nondestructive method for continuously monitoring plant growth

In the past, plant growth generally has been measured using destructive methods. This paper describes a nondestructive technique for continuously monitoring plant growth. The technique provides a means of directly and accurately measuring plant growth over both short and long time intervals. Application of this technique to the direct measurement of plant growth rates is illustrated using corn (Zea mays L.) as an example.

NASA Program CELSS↗

Effects of artificial enclosure of young lettuce leaves on tipburn incidence and leaf calcium concentration

The young developing leaves of 20-day-old lettuce plants (Lactuca sativa L. 'Buttercrunch') were enclosed by aluminized polyethylene sheaths to decrease transpiration and reduce Ca transport. The plants were grown in recirculating solution culture system using a modified half-strength Hoagland's solution under cool-white fluorescent lamps with a photosynthetic photon flux of 350 micromoles s-1 m-2 in a 16:8-hr (light:dark) period. Air temperature and humidity were 20 degrees C and 65%, respectively. After 4 days of enclosure, 53% of the inner leavers (leaves one to 3 cm in length) were tipburned. After the same period, less than 1% of the inner leaves on control plants were tipburned. The concentration of Ca in enclosed inner leaves was 0.63 mg g-1 dry weight, compared to 1.48 mg g-1 dry weight in inner leaves that were not enclosed. The Ca concentration in transpiring outer leaves of all plants was 9.9 mg g-1 dry weight. The Mg concentration in enclosed inner leaves was 2.25 mg g-1 dry weight, compared to 2.34 mg g-1 dry weight in inner leaves that were not enclosed. This research documents that enclosures of leaves at the growing point, as would occur with normal head development, is sufficient to create a limiting concentration of Ca in the enclosed tissue and encourage tipburn development.

Non-NASA Center↗

Modeling the effects of light, carbon dioxide, and temperature on the growth of potato

This study examined the effects of light, temperature and carbon dioxide on the growth of potato (Solanum tuberosum L.) in a controlled environment in order to ascertain the best growing conditions for potato in life support systems in space. 'Norland' and 'Russet Burbank' were grown in 6-L pots of peat-vermiculite for 56 d in growth chambers at the University of Wisconsin Biotron. Environmental factor levels included continuous light (24-h photoperiod) at 250, 400, and 550 micromoles m-2 s-1 PPF; constant temperature at 16, 20, and 24 degrees C; and CO2 at approximately 400, 1000, and 1600 microliters L-1. Separate effects analysis and ridge analysis provided a means to examine the effects of individual environmental factors and to determine combinations of factors that are expected to give the best increases in yields over the central design point. The response surface of Norland indicated that tuber yields were highest with moderately low temperature (18.7 degrees C), low CO2 (400 microliters L-1) and high light (550 micromoles m-2 s-1 PPF). These conditions also favored shorter stem growth. Russet Burbank tuber yields were highest at moderately low temperature (17.5 degrees C), high CO2 (1600 microliters L-1) and medium analyses will be used to project the most efficient conditions for growth of potatoes in closed ecological life support systems (CELSS) in space colonies.

NASA Discipline Life Support Systems↗

Proton gradients in intact cyanobacteria

The internal pH values of two unicellular cyanobacterial strains were determined with electron spin resonance probes, over an external pH range of 6 to 9, in the light and in the dark. The slow growing, thylakoid-lacking Gloeobacter violaceus was found to have a low capacity for maintaining a constant internal pH. The distribution pattern of weak acid and amine nitroxide spin probes across the cell membranes of this organism, in the light and in the dark, was consistent with the assumption that it contains a single intracellular compartment. At an external pH of 7.0, intracellular pH was 6.8 in the dark and 7.2 in the light. The cells of Agmenellum quadruplicatum, a marine species, were found to contain two separate compartments; in the dark, the pH of the cytoplasmic and the intrathylakoid spaces were calculated to be 7.2 and 5.5, respectively. Upon illumination, the former increased and the latter decreased by about 0.5 pH units.

Non-NASA Center↗

Microbial biomass and productivity in seagrass beds

Different methods for measuring the rates of processes mediated by bacteria in sediments and the rates of bacterial cell production have been compared. In addition, net production of the seagrass Zostera capricorni and bacterial production have been compared and some interrelationships with the nitrogen cycle discussed. Seagrass productivity was estimated by measuring the plastochrone interval using a leaf stapling technique. The average productivity over four seasons was 1.28 +/- 0.28 g C m-2 day-1 (mean +/- standard deviation, n = 4). Bacterial productivity was measured five times throughout a year using the rate of tritiated thymidine incorporated into DNA. Average values were 33 +/- 12 mg C m-2 day-1 for sediment and 23 +/- 4 for water column (n = 5). Spatial variability between samples was greater than seasonal variation for both seagrass productivity and bacterial productivity. On one occasion, bacterial productivity was measured using the rate of 32P incorporated into phospholipid. The values were comparable to those obtained with tritiated thymidine. The rate of sulfate reduction was 10 mmol SO4(-2) m-2 day-1. The rate of methanogenesis was low, being 5.6 mg CH4 produced m-2 day-1. A comparison of C flux measured using rates of sulfate reduction and DNA synthesis indicated that anaerobic processes were predominant in these sediments. An analysis of microbial biomass and community structure, using techniques of phospholipid analysis, showed that bacteria were predominant members of the microbial biomass and that of these, strictly anaerobic bacteria were the main components. Ammonia concentration in interstitial water varied from 23 to 71 micromoles. Estimates of the amount of ammonia required by seagrass showed that the ammonia would turn over about once per day. Rapid recycling of nitrogen by bacteria and bacterial grazers is probably important.

NASA Program CELSS↗

Tomato responses to ammonium and nitrate nutrition under controlled root-zone pH

Tomato (Lycopersicon esculentum L. Mill. 'Vendor') plants were grown for 21 days in flowing solution culture with N supplied as either 1.0 mM NO3- or 1.0 mM NH4+. Acidity in the solutions was automatically maintained at pH 6.0. Accumulation and distribution of dry matter and total N and net photosynthetic rate were not affected by source of N. Thus, when rhizosphere acidity was controlled at pH 6.0 during uptake, either NO3- or NH4+ can be used efficiently by tomato. Uptake of K+ and Ca2+ were not altered by N source, but uptake of Mg2+ was reduced in NH4(+)-fed plants. This indicates that uptake of Mg2+ was regulated at least partially by ionic balance within the plant.

NASA Program CELSS↗