A multiple-trait breeding program for improving the symbiosis for N2 fixation betweenMedicago sativa L. andRhizobium meliloti

Summary The goal of breeding alfalfa for increased N₂ fixation potential is addressed. A chronological progression of breeding, physiological, microbiological, and plant pathological research is described. Studies describing the interrelationships among plant morphological, plant physiological, andRhizobium effectiveness traits are summarized. It was concluded that N₂ fixation in alfalfa is affected by coordinated responses among many physiological and biochemical traits. The simultaneous improvement of many factors in the symbiosis requires a comprehensive multiple-step breeding program. The current program includes selection in the glasshouse for seedling vigor,Rhizobium preference, shoot growth, nodule mass, root growth, nitrogenase (as measured by acetylene reduction), and nodule enzyme activity. The inclusion of additional selection traits is anticipated. Field evaluations of N₂ fixation potential of alfalfa populations are made with¹⁵N isotope dilution techniques. Plant germplasm sources used in the breeding program include several heterogeneous populations which have good combining ability and pest resistance when they are intercrossed. Significant progress has been made in achieving the goal of breeding alfalfa for improved N₂ fixation.     

Understanding crop physiology to select breeding targets and improve crop management under increasing atmospheric CO2 concentrations

The present overview paper reviews knowledge on plant metabolism under elevated atmospheric CO₂ concentrations (e[CO₂]) with regard to underpinning options for the management of crop production systems and the selection of crop traits beneficial for future conditions.Better understanding of intra-specific variability in responses to e[CO₂] is of great importance to breed or select best possible genotypes for future conditions. Yield increases per 100 μL L⁻¹ increase in [CO₂] varied between none and over 30% among varieties of important crops. Carbon source–sink relationships are believed to play a major role in determining the ability of a plant to utilise e[CO₂] and avoid downward acclimation of photosynthesis upon prolonged e[CO₂] exposure. Corresponding traits (e.g. tillering capacity, stem carbohydrate storage capacity, or seed size and numbers) are currently under investigation in Free Air Carbon dioxide Enrichment (FACE) facilities, such as AGFACE (Australian Grains FACE).The stimulatory effect of e[CO₂] on plant growth is dependent on adequate nutrient supply. For example, N concentrations in plant tissues generally decrease under e[CO₂], which in leaves is commonly related to a decrease in Rubisco concentration and activity, and therefore linked to photosynthetic downward acclimation. This effect is also of direct concern for food production where decreased N and protein content can have negative effects on product quality (e.g. grain protein). Plant nutrient metabolism appears to adjust to a new physiological equilibrium under e[CO₂] which limits the extent to which nutrient application can ameliorate the situation. What the control points are for an adjustment of plant N metabolism is unclear. Rubisco metabolism in leaves, N assimilation, N translocation or N uptake are all potential key steps that may be inhibited or downregulated under e[CO₂]. To achieve the best possible growth response whilst maintaining product quality, it is important to understand plant nutrient metabolism under e[CO₂].Comparatively little is known about mechanisms of potential changes in plant stress tolerance under e[CO₂]. Defence metabolites such as antioxidants are, in part, directly linked to primary carbohydrate mechanism and so potentially impacted by e[CO₂]. It is unknown whether photoprotective and antioxidative defence systems, key to plant stress tolerance, will be affected, and if so, whether the response will be strengthened or weakened by e[CO₂]. Better understanding of underlying principles is particularly important because it is virtually impossible to test all possible stress factor combinations with e[CO₂] in realistic field settings.     

Cultivar and cultivar x environment effects on the development of callus and polyhaploid plants from anther cultures of wheat

Summary Plants of three common wheat (Triticum aestivum L. em. Thell) cultivars and one randomly selected doubled-haploid line derived by anther culture from each of the three cultivars were each grown in three environments, a field environment, a greenhouse environment, and a growth chamber environment. Anthers containing largely miduninucleate to late uninucleate microspores were cultured and calli were induced to regenerate plants in order to assess the effects of cultivar, cultivar family (cultivar and corresponding doubled-haploid derivative), anther-donor plant environment, and cultivar X environment interaction on androgenic responses. Large differences in response were observed among cultivars as well as between cultivars and doubled-haploids. Differences between cultivar and doubled-haploid within cultivar family usually resulted from higher frequency of response in the cultivar, contrary to the hypothesis that anther culture per se constitutes a general selective device for superior androgenic responses. Also, in a second experiment, anther callusing frequency was greater in the cultivar Kitt than in any of five unique doubled-haploid lines derived from Kitt. Significant effects were also observed in the first experiment for the interactions of cultivar family X environment as well as doubled-haploid vs. cultivar X environment, although the effect of environment itself was less significant than these interactions.Contribution from the USDA, SEA, AR, Beltsville, Md, and the Department of Agronomy, University of Maryland, College Park, Md, as scientific article No. A-3413, contribution No. 6486     

A numerical simulation of plant growth

A simulation model has been formulated to calculate the growth of plants as a function of the environmental factors: light intensity, temperature and water. The environmental factors are derived from conventional meteorological variables. A plant is defined in terms of its physiological requirements for light, heat and water. Observed plant responses are used to develop functional relationships between the rate of biomass accumulation and the environmental factors. Simulations of corn growth based upon numerical integrations of observed meteorological variables are compared with measured crop yields.Presented at the Seventh International Biometeorological Congress, 17–23 August 1975, College Park, Maryland, USA.     

Isozyme marker loci associated with cold tolerance and maturity in maize

Summary Two maize (Zea mays L.) populations, AS1(S) and ECR-A, were evaluated for allozyme frequency changes associated with selection for improved seedling emergence, early season vigor and early maturity. Eleven marker loci were examined and four loci were used for indirect selection in an attempt to modify cold tolerance and maturity. Allozyme-selected divergent subpopulations were produced by compositing selected S₁ progeny from cycle one (C1) of AS1(S) and from C2 of ECR-A. These subpopulations and S₁ generations from all cycles resulting from phenotypic selection, ECR-A C1 through C7 and AS1(S) CO through C6, were tested in cold tolerance and agronomic performance trials over five environments in 1986. Seedling emergence and seedling dry weight did not improve with phenotypic selection in ECR-A, while plant height, ear height, grain yield, grain moisture, days to mid-silk and days to mid-pollen were reduced significantly. Contrasts between divergent allozyme-selected subpopulations from ECR-A were significant for grain moisture and mid-pollen date. For AS1(S), seeding emergence increased, while plant and ear height decreased with phenotypic selection. Contrasts between allozyme-selected subpopulations were significant for plant and ear height. Changes associated with marker-based selection for AS1(S) were not in the same direction as with phenotypic selection. Selection for favorable allozyme genotypes may be effective in changing certain traits in populations that have been modified by direct selection, however results may not be predictable.Contribution from the Department of Agronomy, Wisconsin Agric. Exp. Stn., Madison, WI. Part of a thesis submitted by the senior author in partial fulfillment of the requirements for a Ph. D. received June, 1987. Research supported by the College of Agric. and Life Sci., University of Wisconsin-Madison, Dekalb-Pfizer Genetics, Garst Seed Company, and Pioneer Hi-Bred.     

Nitrogen fixation by subclover associations fertilized with sulfur

Summary The effect of S fertilization on symbiotic N₂ fixation was measured with the¹⁵N technique and the N difference method in a lysimeter study using Josephine loam (Typic Haploxurults). Nitrogen fixation by subclover (Trifolium subterraneum L.) was strongly enhanced by added S. The association of soft chess (Bromus mollis L.) or filaree (Erodium botrys (Cav.) Bertol.) with subclover increased the percentage of N in subclover that was fixed, with the results that N₂ fixation was increased beyond that due to the mere increase in subclover biomass. Nitrogen fixation estimates by¹⁵N dilution and N difference methods were highly correlated (r²=0.97), and S fertilizer did not result in any significant differences in N₂-fixation estimation by the two methods. Both methods were useful in distinguishing between soil N uptake and N₂ fixation where S applications produced highly significant increases in both uptake and fixation. Application of sulfur fertilizers to much annual rangeland has the potential to increase pasture productivity through enhanced N₂ fixation. Contribution of the University of California Hopland Field Station and Department of Agronomy and Range Science, Univ. of California, Davis, CA 95616.     

Demand and supply of N in seed production of soybean (<Emphasis Type="Italic">Glycine max</Emphasis>) at different N fertilization levels after flowering

Nitrogen (N) has been suggested as a determinant of seed production especially in species with high seed N content. Assuming that seed yield was determined as the balance between N demand and supply for seed production, we studied the effect of N fertilization after flowering on soybean (Glycine max L. Merr.) yield. Seed N concentration was nearly constant irrespective of N fertilization, indicating that seed production was proportional to the amount of N available for seed growth. N demand for seed production was analyzed as the product of seed number, the rate of N filling in individual seeds, and the length of the reproductive period. N fertilization increased seed number and the reproductive period, but did not influence the N filling rate. Seed number was positively correlated with dry mass productivity after flowering. Three N sources were distinguished: mineral N uptake, symbiotic N₂ fixation and N remobilization from vegetative body. N fertilization increased N uptake and N remobilization, but lowered N₂ fixation. We concluded that N availability in the reproductive period determined seed yield directly through increasing N supply for seed growth and indirectly through increasing seed N demand with enhanced plant dry mass productivity.     

Resistance of soil protein depolymerization rates to eight years of elevated CO<Subscript>2</Subscript>, warming,and summer drought in a temperate heathland

Soil N availability for plants and microorganisms depends on the breakdown of soil polymers such as proteins into smaller, assimilable units by microbial extracellular enzymes. Changing climatic conditions are expected to alter protein depolymerization rates over the next decades, and thereby affect the potential for plant productivity. We here tested the effect of increased CO₂ concentration, temperature, and drought frequency on gross rates of protein depolymerization, N mineralization, microbial amino acid and ammonium uptake using ¹⁵N pool dilution assays. Soils were sampled in fall 2013 from the multifactorial climate change experiment CLIMAITE that simulates increased CO₂ concentration, temperature, and drought frequency in a fully factorial design in a temperate heathland. Eight years after treatment initiation, we found no significant effect of any climate manipulation treatment, alone or in combination, on protein depolymerization rates. Nitrogen mineralization, amino acid and ammonium uptake showed no significant individual treatment effects, but significant interactive effects of warming and drought. Combined effects of all three treatments were not significant for any of the measured parameters. Our findings therefore do not suggest an accelerated release of amino acids from soil proteins in a future climate at this site that could sustain higher plant productivity.     

Regulation of Nitrogen Metabolism,Photosynthetic Activity,and Yield Attributes of Spring Wheat by Nitrogen Fertilizer in the Semi-arid Loess Plateau Region

It is critical for spring wheat (Triticum aestivum L.) production in the semi-arid Loess Plateau to understand the impact of nitrogen (N) fertilizer on changes in N metabolism, photosynthetic parameters, and their relationship with grain yield and quality. The photosynthetic capacity of flag leaves, dry matter accumulation, and N metabolite enzyme activities from anthesis to maturity were studied on a long-term fertilization trial under different N rates [0 kg ha^?1(N1), 52.5 kg ha^?1 (N2), 105 kg ha^?1 (N3), 157.5 kg ha^?1 (N4), and 210 kg ha^?1 (N5)]. It was observed that N3 produced optimum total dry matter (5407 kg ha^?1), 1000 grain weight (39.7 g), grain yield (2.64 t ha^?1), and protein content (13.97%). Our results showed that N fertilization significantly increased photosynthetic parameters and N metabolite enzymes at all growth stages. Nitrogen harvest index, partial productivity factor, agronomic recovery efficiency, and nitrogen agronomic efficiency were decreased with increased N. Higher N rates (N3–N5) maintained higher photosynthetic capacity and dry matter accumulation and lower intercellular CO₂ content. The N supply influenced NUE by improving photosynthetic properties. The N3 produced highest chlorophyll content, photosynthetic rate, stomatal conductance and transpiration rate, grain yield, grain protein, dry matter, grains weight, and N metabolite enzyme activities compared to the other rates (N1, N2, N4, and N5). Therefore, increasing N rates beyond the optimum quantity only promotes vegetative development and results in lower yields.

     

Concurrent activation of OsAMT1;2 and OsGOGAT1 in rice leads to enhanced nitrogen use efficiency under nitrogen limitation

Nitrogen (N) is a major factor for plant development and productivity. However, the application of nitrogenous fertilizers generates environmental and economic problems. To cope with the increasing global food demand, the development of rice varieties with high nitrogen use efficiency (NUE) is indispensable for reducing environmental issues and achieving sustainable agriculture. Here, we report that the concomitant activation of the rice (Oryza sativa) Ammonium transporter 1;2 (OsAMT1;2) and Glutamate synthetase 1 (OsGOGAT1) genes leads to increased tolerance to nitrogen limitation and to better ammonium uptake and N remobilization at the whole plant level. We show that the double activation of OsAMT1;2 and OsGOGAT1 increases plant performance in agriculture, providing better N grain filling without yield penalty under paddy field conditions, as well as better grain yield and N content when plants are grown under N llimitations in field conditions. Combining OsAMT1;2 and OsGOGAT1 activation provides a good breeding strategy for improving plant growth, nitrogen use efficiency and grain productivity, especially under nitrogen limitation, through the enhancement of both nitrogen uptake and assimilation.     

Protein differential expression in the elongating cotton (Gossypium hirsutum L.) fiber under nitrogen stress

Nitrogen (N) is an essential macronutrient and an important factor limiting agricultural productivity. N deficient or excess conditions often occur during the cotton growth season and incorrect N application may affect cotton fiber yield and quality. Here, the influence of N stress on the cotton fiber proteome was investigated by two-dimensional gel electrophoresis and mass spectrometry. The results indicated that N application rate affects nitrogen accumulation in fiber cells and fiber length. The proteins differentially expressed during N stress were mainly related to plant carbohydrate metabolism, cell wall component synthesis and transportation, protein/amino acid metabolism, antioxidation and hormone metabolism. The most abundant proteins were C metabolism-related. Ten days post anthesis is a critical time for fiber cells to perceive environmental stress and most proteins were suppressed in both N deficient and N excess conditions at this sampling stage. However, several N metabolism proteins were increased to enhance N stress tolerance. Excess N may suppress carbohydrate/energy metabolism in early fiber development much like N deficiency. These results have identified some interesting proteins that can be further analyzed to elucidate the molecular mechanisms of N tolerance.     

Mapping of QTL associated with nitrogen storage and remobilization in barley (Hordeum vulgare L.) leaves

Nitrogen uptake and metabolism are central for vegetative and reproductive plant growth. This is reflected by the fact that nitrogen can be remobilized and reused within a plant, and this process is crucial for yield in most annual crops. A population of 146 recombinant inbred barley lines (F(8) and F(9) plants, grown in 2000 and 2001), derived from a cross between two varieties differing markedly in grain protein concentration, was used to compare the location of QTL associated with nitrogen uptake, storage and remobilization in flag leaves relative to QTL controlling developmental parameters and grain protein accumulation. Overlaps of support intervals for such QTL were found on several chromosomes, with chromosomes 3 and 6 being especially important. For QTL on these chromosomes, alleles associated with inefficient N remobilization were associated with depressed yield and higher levels of total or soluble organic nitrogen during grain filling and vice versa; therefore, genes directly involved in N recycling or genes regulating N recycling may be located on these chromosomes. Interestingly, the most prominent QTL for grain protein concentration (on chromosome 6) did not co-localize with QTL for nitrogen remobilization. However, QTL peaks for nitrate and soluble organic nitrogen were detected at this locus for plants grown in 2001 (but not in 2000). For these, alleles associated with low grain protein concentration were associated with higher soluble nitrogen levels in leaves during grain filling; therefore, gene(s) found at this locus might influence the nitrogen sink strength of developing barley grains.     

Protein accumulation potential in barley seeds as affected by soil- and peduncle-applied N and peduncle-applied plant growth regulators

Although much investigated, the factors constraining cereal grain protein accumulation are not well understood. As a result of the development of a new technique, new approaches to this question are now possible. A peduncle perfusion system was used to deliver a range of plant growth regulators (PGRs) and/or N solutions into barley (Hordeum vulgare) plants during the grain-filling period. The perfusion technique floods the peduncle interior with a treatment solution for periods of weeks to months, allowing the plant to take up administered substances from the perfused solution. The objectives of the present work were to determine: (1) whether some PGRs could alter the overall pattern of N allocation within barley plants, perhaps leading to higher protein accumulation in the seeds, (2) whether the addition of N through the peduncle could increase the seed N concentration even when the concentration of N in the rooting medium was high, and (3) whether or not PGR-stimulated elevations in grain protein levels and peduncle-added N increases in grain protein levels were additive. Three experiments were conducted to determine the physiological effects of (1) peduncle-administered PGRs (2) combinations of soil- and peduncle-applied N and (3) selected combinations of soil- and peduncle-administered N, and peduncle-applied PGRs on photosynthetic rate, dry matter partitioning and N accumulation of barley plants during grain filling. The first experiment tested four PGRs: abscisic acid (ABA), kinetin, gibberellic acid (GA₃), and 2,4-dichlorophenoxy acetic acid (2,4-D) each at three concentrations. The second experiment tested three levels of soil N (NH₄NO₃) fertility, and two concentrations of peduncle-added N (urea). The third experiment tested four PGRs: ABA, kinetin, GA₃, and 2,4-D with two soil N concentrations and two concentrations of peduncle-added N. ABA and 2,4-D decreased total seed weight of the perfused spike. The addition of peduncle-perfused N increased seed protein concentration and content under conditions of high soil N fertility, suggesting that seed protein accumulation is more limited by the ability of roots to take up N from the soil than by the seed to take up N from the rest of the plant. The effects of the PGRs on N allocation among plant parts varied with the amount of N available to the plant. Because it resulted in less protein stored in the flag leaf and more in the seeds, GA₃ perfusion caused an overall change in the allocation of N among plant parts. Peduncle perfusion of kinetin and ABA affected some aspects of photosynthetic physiology.     

Changes in C–N metabolism under elevated CO2 and temperature in Indian mustard (Brassica juncea L.): an adaptation strategy under climate change scenario

The present study was performed to investigate the possible role of carbon (C) and nitrogen (N) metabolism in adaptation of Indian mustard (Brassica juncea L.) growing under ambient (370 ± 15 ppm) and elevated CO₂ (700 ± 15 ppm), and jointly in elevated CO₂ and temperature (30/22 °C for day/night). The key enzymes responsible for C–N metabolism were studied in different samples of Brassica juncea L. collected from ambient (AMB), elevated (ELE) and ELExT growth conditions. Total percent amount of C and N in leaves were particularly estimated to establish a clear understanding of aforesaid metabolism in plant adaptation. Furthermore, key morphological and physiological parameters such as plant height, leaf area index, dry biomass, net photosynthetic rate, stomatal conductance, transpiration, total protein and chlorophyll contents were also studied in relation to C/N metabolism. The results indicated that the C-metabolizing enzymes, such as (ribulose-1,5-bisphosphate carboxylase/oxygenase, phosphoenolpyruvate carboxylase, malate dehydrogenase, NAD-malic enzyme, NADP-malic enzyme and citrate synthase) and the N-metabolizing enzymes, such as (aspartate amino transferase, glutamine synthetase, nitrate reductase and nitrite reductase) showed significantly (P < 0.05) higher activities along with the aforesaid physiological and biochemical parameters in order of ELE > ELExT > AMB growth conditions. This is also evident by significant (P < 0.05) increase in percent contents of C and N in leaves as per said order. These findings suggested that improved performance of C–N metabolism could be a possible approach for CO₂ assimilation and adaptation in Brassica juncea L. against elevated CO₂ and temperature prevailing in climate change scenarios.     

推迟拔节水对小麦氮素积累与分配和硝态氮运移的影响

摘要：2007—2008年度以高产冬小麦品种济麦22为材料,设置2个拔节水灌溉时期,为拔节期和拔节后10 d;3个目标相对含水量,灌水后0~140 cm土层土壤相对含水量分别达到65%、75%、80%,以W1、W2、W3表示拔节期灌水处理,DW1、DW2、DW3表示拔节后10 d灌水处理;开花期均灌水至0~140 cm土层土壤相对含水量为70%,研究推迟拔节水对小麦氮素积累与分配和硝态氮运移的影响。结果表明：（1）W2和DW2处理有利于提高0~60 cm土层土壤硝态氮含量,促进籽粒氮素积累;营养器官贮藏氮素向籽粒的转运量、籽粒产量和氮肥偏生产力分别高于W1和DW1,与W3和DW3处理无显著差异;开花后植株氮素积累量、籽粒蛋白质含量和水分利用效率分别高于W3和DW3,是拔节期和拔节后10 d灌水的最优处理。（2）W2和DW2处理比较,DW2成熟期100~140 cm土层硝态氮残留量低于W2,籽粒产量、籽粒蛋白质含量、氮素吸收效率、氮肥偏生产力和水分利用效率均显著高于W2,是本试验条件下的最佳灌水方案。2008—2009生长季试验各处理变化趋势同2007—2008年度。     

Origins and metabolism of formate in higher plants

Formate, a simple one-carbon compound, is readily metabolized in plant tissues. In greening potato tubers, similar to some procaryotes, formate is directly synthesized via a ferredoxin-dependent fixation of CO₂, serving as the main precursor for carbon skeletons in biosynthetic pathways. In other plant species and tissues, formate appears as a side-product of photorespiration and of fermentation pathways, but possibly also as a product of direct CO₂ reduction in chloroplasts. Formate metabolism is closely related to serine synthesis and to all subsequent reactions originating from serine. Formate may have a role in biosynthesis of numerous compounds, in energetic metabolism and in signal transduction pathways related to stress response. This review summarizes the current state of formate research, physiological/biochemical and molecular aspects.