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The elite haplotype OsGATA8 -H coordinates nitrogen uptake and productive tiller formation in rice

Excessive nitrogen promotes the formation of nonproductive tillers in rice, which decreases nitrogen use efficiency (NUE). Developing high-NUE rice cultivars through balancing nitrogen uptake and the formation of productive tillers remains a long-standing challenge, yet how these two processes are coordinated in rice remains elusive. Here we identify the transcription factor OsGATA8 as a key coordinator of nitrogen uptake and tiller formation in rice. OsGATA8 negatively regulates nitrogen uptake by repressing transcription of the ammonium transporter gene OsAMT3.2. Meanwhile, it promotes tiller formation by repressing the transcription of OsTCP19, a negative modulator of tillering. We identify OsGATA8-H as a high-NUE haplotype with enhanced nitrogen uptake and a higher proportion of productive tillers. The geographical distribution of OsGATA8-H and its frequency change in historical accessions suggest its adaption to the fertile soil. Overall, this study provides molecular and evolutionary insights into the regulation of NUE and facilitates the breeding of rice cultivars with higher NUE.

59 BASIC BIOLOGICAL SCIENCES

Comparing field and lab quantitative stable isotope probing for nitrogen assimilation in soil microbes

ABSTRACT Soil microbial communities play crucial roles in nutrient cycling and can help retain nitrogen in agricultural soils. Quantitative stable isotope probing (qSIP) is a useful method for investigating taxon-specific microbial growth and utilization of specific nutrients, such as nitrogen (N). Typically, qSIP is performed in a highly controlled lab setting, so the field relevance of lab qSIP studies remains unknown. We conducted and compared tandem lab and field qSIP to quantify the assimilation of 15 N by maize-associated soil prokaryotic communities at two agricultural sites. Here, we show that field qSIP with 15 N can be used to measure taxon-specific microbial N assimilation. Relative 15 N assimilation rates were generally lower in the field, and the magnitude of this difference varied by site. Rates differed by method (lab vs field) for 19% of the top N assimilating genera. The field and lab measures were more comparable when relative assimilation rates were weighted by relative abundance to estimate the proportion of N assimilated by each genus with only ~10% of taxa differing by method. Of those that differed, the taxa consistently higher in the lab were inclined to have opportunistic lifestyle strategies, whereas those higher in the field had niches reliant on plant roots or in-tact soil structure (biofilms, mycelia). This study demonstrates that 15 N-qSIP can be successfully performed using field-incubated soils to identify microbial allies in N retention and highlights the strengths and limitations of field and lab qSIP approaches. IMPORTANCE Soil microbes are responsible for critical biogeochemical processes in natural and agricultural ecosystems. Despite their importance, the functional traits of most soil organisms remain woefully under-characterized, limiting our ability to understand how microbial populations influence the transformation of elements such as nitrogen (N) in soil. Quantitative stable isotope probing (qSIP) is a powerful tool to measure the traits of individual taxa. This method has rarely been applied in the field or with 15 N to measure nitrogen assimilation. In this study, we measured genus-specific microbial nitrogen assimilation in two agricultural soils and compared field and lab 15 N qSIP methods. Our results identify taxa important for nitrogen assimilation in agricultural soils, shed light on the field relevance of lab qSIP studies, and provide guidance for the future application of qSIP to measure microbial traits in the field.

Reed, Kinsey (ORCID:0000000155178664)

Phylogenomic discovery and engineering of nitrogen fixation into the bioenergy woody crop poplar

Biological nitrogen fixation (BNF) is a key process enabling plants in specific lineages to convert atmospheric dinitrogen (N₂) into bioavailable ammonia through symbioses with diazotrophic microbes. Expanding this capability beyond native nitrogen-fixing clades into non-nodulating crops would reduce synthetic fertilizer use, lowering energy inputs and environmental impacts in agriculture. Supported by DOE Funding Award DE-SC0018247, the NitFix project advanced foundational knowledge required to engineer root-nodule symbioses in new host species. The team generated the most comprehensive phylogenomic analysis to date of all known nodulating lineages, resolving the evolutionary history of nitrogen-fixing symbiosis and identifying core gene suites retained across nodulating taxa. Through multimodal genomics, transcriptomics, and functional analyses in Medicago truncatula and related species, the project mapped regulatory networks underlying nodule organogenesis, bacterial infection, and nitrogen-fixation efficiency. Key discoveries include the identification of conserved signaling modules for rhizobial recognition, transcription factors controlling nodule differentiation, and metabolic pathways integrating fixed nitrogen into plant growth. The project also developed enabling tools—including optimized transformation pipelines, gene-editing workflows, and imaging-based phenotyping—to accelerate engineering efforts in emerging models. Together, these results refine the mechanistic framework of symbiotic nitrogen fixation and highlight transferable components essential for rewiring these traits into non-nodulating crops.

59 BASIC BIOLOGICAL SCIENCES

Plasma polymerization of an ethylene-nitrogen gas mixture

A procedure has been developed whereby nitrogen can be incorporated into an organic film from an ethylene-nitrogen gas mixture using an internal electrode capacitively coupled radio frequency reactor. The presence of nitrogen has been shown directly by infrared transmittance spectra and electron spectroscopic chemical analysis data, and further indirect evidence was provided by dielectric measurements and by the reverse osmosis properties of the film. Preparation of a nitrogen containing film did not require vapor from an organic nitrogen containing liquid monomer. Some control over the bonding and stoichiometry of the polymer film was provided by the added degree of freedom of the nitrogen partial pressure in the gas mixture. This new parameter strongly affected the dielectric properties of the plasma polymerized film and could affect the reverse osmosis behavior.

Hudis, M.

Evaluation of a spacecraft nitrogen generator

A research and development program was successfully completed towards the development of a method of generating nitrogen for cabin leakage makeup aboard space vehicles. The nitrogen generation concept used liquid hydrazine as the stored form of nitrogen. This reduced tankage and expendables weight associated with high pressure gaseous and cryogenic liquid nitrogen storage. The hydrazine was catalytically dissociated to yield a mixture of nitrogen and hydrogen. The latter was separated to provide the makeup nitrogen. The hydrogen will be used in the reduction of metabolic carbon dioxide.

Marshall, R. D.

Effect of oxygen and nitrogen interactions on friction of single-crystal silicon carbide

Friction studies were conducted with single-crystal silicon carbide contacting silicon carbide and titanium after having been exposed to oxygen and nitrogen in various forms. After they had been sputter cleaned, the surfaces were (1) exposed to gaseous oxygen and nitrogen (adsorption), (2) ion bombarded with oxygen and nitrogen, or (3) reacted with oxygen (SiC only). Auger emission spectroscopy was used to determine the presence of oxygen and nitrogen. The results indicate that the surfaces of silicon carbide with reacted and ion-bombarded oxygen ions give higher coefficients of friction than do argon sputter-cleaned surfaces. The effects of oxygen on friction may be related to the relative chemical, thermodynamic properties of silicon, carbon, and titanium for oxygen. The adsorbed films of oxygen, nitrogen, and mixed gases of oxygen and nitrogen on sputter-cleaned, oxygen-ion bombarded, and oxygen-reacted surfaces generally reduce friction. Adsorption to silicon carbide is relatively weak.

Miyoshi, K.

Nitrogen abundances and isotopic compositions in lunar samples

Isotopic analyses were carried out on soil separates as well as on bulk samples. Analyses of nitrogen fractions obtained by step-wise heating of the separates were also conducted. It was also attempted to obtain a value for the isotopic composition of indigenous lunar nitrogen from analyses of igneous rock samples. Several breccias were also analyzed for their nitrogen isotope ratios. The significance of the obtained results is discussed. Several lines of evidence point to the conclusion that the isotope ratio of the nitrogen being implanted into the lunar regolith has increased by some 15% over a period of at least 450 million years and possibly as long as 3,700 million years or more. This may be the result of changes in the nitrogen isotope ratio of the solar wind with time, or it may be due to outgassing and subsequent reimplantation of an isotopically light indigenous lunar nitrogen from the lunar interior in the early history of the moon.

Becker, R. H.

Odd nitrogen production by meteoroids

The process by which odd nitrogen species (atomic nitrogen and nitric oxide) are formed during atmospheric entry of meteoroids is analyzed theoretically. An ablating meteoroid is assumed to be a point source of mass with a continuum regime evolving in its wake. The amounts of odd nitrogen species, produced by high-temperature reactions of air in the continuum wake, are calculated by numerical integration of chemical rate equations. Flow properties are assumed to be uniform across the wake, and 29 reactions involving five neutral species and five singly ionized species are considered, as well as vibrational and electron temperature nonequilibrium phenomena. The results, when they are summed over the observed mass, velocity, and entry-angle distribution of meteoroids, provide odd-nitrogen-species annual global production rates as functions of altitude. The peak production of nitric oxide is found to occur at an altitude of about 85 km; atomic nitrogen production peaks at about 95 km. The total annual rate for nitric oxide is 40 million kg; for atomic nitrogen it is 170 million kg.

Park, C.

High-altitude atomic nitrogen densities

Theoretical calculations of the seasonal and diurnal variations of atomic nitrogen are compared with measurements made by the open source neutral mass spectrometer on the AE-C satellite. With the simultaneous measurements of molecular nitrogen and atomic oxygen densities as input, model calculations of odd nitrogen densities predict the same trends in atomic nitrogen as those observed. From these comparisons it is inferred that horizontal transport significantly reduces the diurnal variation of atomic nitrogen. Estimates are given of the sensitivity of atomic nitrogen densities to variations in the photoelectron flux, the neutral temperatures, and the neutral winds.

Oran, E. S.

Nitrogen isotope systematics of two Apollo 12 soils

Soils 12023 and 12037 were analyzed for nitrogen by step-wise heating. Helium contents, Ar-40/Ar-36 ratios and spallation N-15 ages were determined in addition to nitrogen contents and isotopic compositions. The results for 12023 show it to be a typical lunar soil similar to many Apollo 16 and 17 soils in its nitrogen content and delta N-15 value, with a complex history. Soil 12037 appears to have had a simple, two-stage surface exposure history; one exposure occurring recently for about 10-20 m.y. and the other between about 3 to 4 b.y. ago. The delta N-15 value of nitrogen implanted in the earlier exposure was -125%. This nitrogen may reside in the minor components of 12037 such as the KREEP. If it should reside in the basaltic component of 12037, however, then the variation in the delta N-15 of implanted nitrogen with time probably has been more complex than previously considered.

Becker, R. H.

Depressurization and two-phase flow of water containing high levels of dissolved nitrogen gas

Depressurization of water containing various concentrations of dissolved nitrogen gas was studied. In a nonflow depressurization experiment, water with very high nitrogen content was depressurized at rates from 0.09 to 0.50 MPa per second and a metastable behavior which was a strong function of the depressurization rate was observed. Flow experiments were performed in an axisymmetric, converging diverging nozzle, a two dimensional, converging nozzle with glass sidewalls, and a sharp edge orifice. The converging diverging nozzle exhibited choked flow behavior even at nitrogen concentration levels as low as 4 percent of the saturation level. The flow rates were independent of concentration level. Flow in the two dimensional, converging, visual nozzle appeared to have a sufficient pressure drop at the throat to cause nitrogen to come out of solution, but choking occurred further downstream. The orifice flow motion pictures showed considerable oscillation downstream of the orifice and parallel to the flow. Nitrogen bubbles appeared in the flow at back pressures as high as 3.28 MPa, and the level at which bubbles were no longer visible was a function of nitrogen concentration.

Simoneau, R. J.

Preprototype nitrogen supply subsystem development

A nitrogen supply subsystem based on the dissociation of hydrazine into a mixture of hydrogen and nitrogen is developed. The latter is separated to provide makeup nitrogen to control the composition of spacecraft atmospheres. Specific hardware developments resulted in the design and fabrication of a nominal 3.6 kg/d nitrogen generation module. The design integrates a hydrazine catalytic dissociator, three ammonia dissociation stages and four hydrogen separation stages into a 33 kg, 14 cu dm module. A technique was devised to alternate the ammonia dissociation and hydrogen separation stages to give high nitrogen purity in the end product stream. Tests show the product stream to contain less than 0.5 percent hydrogen and 10 parts per million ammonia. The design and development of a test stand for the nitrogen generation module and a series of tests which verified its operation and performance capability are described.

Heppner, D. B.

Tribological characteristics of nitrogen (N+) implanted iron

The effect of implantation of nitrogen ions (1.5 MeV) on the friction and wear characteristics of pure ion sliding against M-50 steel (unimplanted) was studied in a pin-on-disk sliding friction apparatus. Test conditions included room temperature (25 C), a dry air atmosphere, a load of 1/2 kg (4.9 N), sliding velocities of 0.043 to 0.078 m/sec (15 to 25 rpm), a pure hydrocarbon lubricant (n-hexadecane), or a U.S.P. mineral oil and nitrogen ion implantation doses of 5x10 to the 15th power and 5x10 to the 17th power ions/sq cm. No differences in wear rates were observed in the low dose experiments. In the high dose experiments, small reductions in initial (40 percent) and steady state (20 percent) wear rates were observed for nitrogen implanted iron riders as compared with unimplanted controls. No differences in average friction coefficients were noted for either dose. Auger electron spectroscopy combined with argon ion bombardment revealed a subsurface Gaussian nitrogen distribution with a maximum concentration of 6 atomic percent at a depth of 0.8 microns. Similar analysis within the wear scar of an implanted rider after 20 microns of wear yielded only background nitrogen concentration. No inward migration of nitrogen ions was observed.

Jones, W. R.

Total nitrogen content of deep sea basalts

An estimate of the total nitrogen content of the earth's mantle, aimed at furnishing a further constraint for earth atmosphere origin and evolution models, was attempted through thermal neutron activation analysis via N-14(n,p)C-14 for the case of deep sea basalt glasses from the East Pacific Rise, the Mid-Atlantic Rift, and the Juan de Fuca Ridge. The increased nitrogen abundance of matrix material from the same samples as the glasses may be due to the incorporation of chemically-bound nitrogen from sea water, rather than dissolved molecular nitrogen. A discussion is presented of factors affecting observed basalt nitrogen content and its interpretation in terms of mantle nitrogen abundance. A 2 ppm N lower limit is estimated for the mantle.

Norris, T. L.

Tribological characteristics of nitrogen (N+) implanted iron

The effect of implantation of nitrogen ions (1.5 MeV) on the friction and wear characteristics of pure ion sliding against M-50 steel (unimplanted) was studied in a pin-on-disk sliding friction apparatus. Test conditions included room temperature (25 C), a dry air atmosphere, a load of 1/2 kg (4.9 N), sliding velocities of 0.043 to 0.078 m/sec (15 to 25 rpm), a pure hydrocarbon lubricant (n-hexadecane), or a U.S.P. mineral oil and nitrogen ion implantation doses of 5x10 to the 15th power and 5x10 to the 17th power ions/sq cm. No differences in wear rates were observed in the low dose experiments. In the high dose experiments, small reductions in initial (40 percent) and steady state (20 percent) wear rates were observed for nitrogen implanted iron riders as compared with unimplanted controls. No differences in average friction coefficients were noted for either dose. Auger electron spectroscopy combined with argon ion bombardment revealed a subsurface Gaussian nitrogen distribution with a maximum concentration of 6 atomic percent at a depth of 0.8 microns. Similar analysis within the wear scar of an implanted rider after 20 microns of wear yielded only background nitrogen concentration. No inward migration of nitrogen ions was observed. Previously announced in STAR as N82-24322

Jones, W. R., Jr.

Nitrogen uptake and utilization by intact plants

The results of experiments support the proposed conceptual model that relates nitrogen uptake activity by plants as a balanced interdependence between the carbon-supplying function of the shoot and the nitrogen-supplying function of the roots. The data are being used to modify a dynamic simulation of plant growth, which presently describes carbon flows through the plant, to describe nitrogen uptake and assimilation within the plant system. Although several models have been proposed to predict nitrogen uptake and partitioning, they emphasize root characteristics affecting nutrient uptake and relay on empirical methods to describe the relationship between nitrogen and carbon flows within the plant. Researchers, on the other hand, propose to continue to attempt a mechanistic solution in which the effects of environment on nitrogen (as well as carbon) assimilation are incorporated through their direct effects on photosynthesis, respiration, and aging processes.

Raper, C. D., Jr.

Nitrogen transformations following tropical forest felling and burning on a volcanic soil

Nitrogen transformations and loss were measured following forest clearing in a relatively fertile tropical forest site. Nitrogen mineralization, nitrification, and amounts of ammonium and nitrate increased substantially in surface soils during the 6 mo following burning, then returned to background levels. The nitrogen content of microbial biomass declined to half its original value 6 mo after clearing and remained low in the cleared sites. Plant uptake of nitrogen was substantial on cleared plots (50 g/sq m), but it accounted for only 18 percent of N-15 label added to field plots. MIcrobial immobilization of N-15 was small relative to that in a cleared temperate site, and measurements of denitrification potentials suggested that relatively little mineralized nitrogen was lost to the atmosphere. Substantial amounts of nitrogen (40-70 g/sq m) were retained as exchangeably bound nitrate deep in the soils of a cleared plot on which revegetation was prevented; this process accounted for 12 percent of the N-15 label added to field plots.

Matson, Pamela A.

Simulation of water vapor condensation on LOX droplet surface using liquid nitrogen

The formation of ice or water layers on liquid oxygen (LOX) droplets in the Space Shuttle Main Engine (SSME) environment was investigated. Formulation of such ice/water layers is indicated by phase-equilibrium considerations under conditions of high partial pressure of water vapor (steam) and low LOX droplet temperature prevailing in the SSME preburner or main chamber. An experimental investigation was begun using liquid nitrogen as a LOX simulant. A monodisperse liquid nitrogen droplet generator was developed which uses an acoustic driver to force the stream of liquid emerging from a capillary tube to break up into a stream of regularly space uniformly sized spherical droplets. The atmospheric pressure liquid nitrogen in the droplet generator reservoir was cooled below its boiling point to prevent two phase flow from occurring in the capillary tube. An existing steam chamber was modified for injection of liquid nitrogen droplets into atmospheric pressure superheated steam. The droplets were imaged using a stroboscopic video system and a laser shadowgraphy system. Several tests were conducted in which liquid nitrogen droplets were injected into the steam chamber. Under conditions of periodic droplet formation, images of 600 micron diameter liquid nitrogen droplets were obtained with the stroboscopic video systems.

Powell, Eugene A.