Variation for Nitrogen Use Efficiency Traits in Wheat Under Contrasting Nitrogen Treatments in South-Eastern Europe

Wheat cultivars differ in their response to nitrogen (N) fertilizer, both in terms of its uptake and utilization. Characterizing this variation is an important step in improving the N use efficiency (NUE) of future cultivars while maximizing production (yield) potential. In this study, we compared the agronomic performance of 48 diverse wheat cultivars released between 1936 and 2016 at low and high N input levels in field conditions to assess the relationship between NUE and its components. Agronomic trait values were significantly lower in the low N treatment, and the cultivars tested showed a significant variation for all traits (apart from the N remobilization efficiency), indicating that response is genotype-dependent, although significant genotype × environment effects were also observed. Overall, we show a varietal improvement in NUE over time of 0.33 and 0.30% year–1 at low and high N, respectively, and propose that this is driven predominantly by varietal selection for increased yield. More complete understanding of the components of these improvements will inform future targeted breeding and selection strategies to support a reduction in fertilizer use while maintaining productivity.

A New Perspective on the Role of Glutamine Synthetase in Nitrogen Remobilization in Wheat (Triticum aestivum L.)

Glutamine synthetase (GS), a key enzyme in plant nitrogen metabolism, is closely related to nitrogen remobilization. However, how GS isoforms participate in nitrogen remobilization remains unclear. Here, the spatiotemporal expression of the TaGS gene family after anthesis was investigated, and the results showed that TaGS1;1 was mainly encoded by TaGS1;1-6A, while the other isozymes were mainly encoded by TaGS localized on the A and D subgenomes. TaGS1;2-4A/4D had the highest expression level, especially in rachis and peduncle. Furthermore, immunofluorescence showed TaGS1;2 was located in the phloem of rachis and peduncle. GUS (β-glucuronidase) staining confirmed that ProTaGS1;2-4A/4D::GUS activity was mainly present in the vascular system of leaves, roots, and petal of Arabidopsis. Ureides, an important transport form of nitrogen, were mainly synthesized in flag leaves and transported to grains through the phloem of peduncle and rachis during grain filling. TaAAH, which encodes the enzyme that degrades ureides to release NH4+, had a higher expression in rachis and peduncle and was synchronized with the increase in NH4+ concentration in phloem, indicating that NH4+ in phloem is from ureide degradation. Taking the above into account, TaGS1;2, which is highly expressed in the phloem of peduncle and rachis, may participate in N remobilization by assimilating NH4+ released from ureide degradation.

Effect of Nitrogen Fertilizer Applications on Nitrogen Remobilization and Grain Yield in Soybean (Glycine max [L] Merril) in the Savelugu-Nanton Municipality in Northern Region of Ghana

Two experiments were conducted in the Savelugu-Nanton Municipality of the Northern Region of Ghana in 2012 and 2013 to assess the effect of N fertilizer on growth, N remobilization and grain yield of three local varieties of soybean (Glycine max [L] Merill). The experiments were a 3 x 4 factorial laid in Randomized Complete Block Design (RCBD) with four replications. Factor A was soybean varieties (Jenguma, Quarshie, Ahotor); Factor B was 0, 15, 30 and 45 kg N ha-1. The experimental fields were planted manually on the flat by drilling and later thinned to 2 plants hill-1 at 0.50 m × 0.10 m with a population of about 400 000 plants ha-1. Growth and yield parameters measured were plant height, nodule number plant-1, nodule dry weight plant-1, percent nodule effectiveness, number of pods plant-1, number of seeds pod-1, 100 seed weight, harvest index and grain yield. The results showed that the control recorded lower figures in all growth parameters. Nitrogen remobilization was also observed in all plots, which indicate that soybean needs greater levels of N during grain filling. Again, N remobilization and soybean yield were highest in the 45 kg N ha-1 treatment compared to the other treatments. However, considering the overall yields, farmers in the study area should be advised to adopt starter N fertilization of soybean for higher yields as the soils are highly degraded in soil fertility.

Partitioning and Translocation of Dry Matter and Nitrogen During Grain Filling in Spring Barley Varieties and Their Roles in Determining Malting Quality

To meet the strict requirements for the malting quality of both grain size and protein content for malting barley, a better understanding of the partitioning and remobilization of dry matter (DM) and nitrogen (N) from individual vegetative organs during grain filling may contribute to adjusting a balance in both quality parameters to satisfy the malting criteria of the brewing industry. A 2-year experiment that included 23 spring malting barley varieties was carried out to determine the DM and N partitioning in different organs at anthesis and maturity and to estimate their remobilization to grains. In contrast to the genetic variation of the 23 barley varieties, year effect was the most important single factor influencing the DM and N accumulation at pre-anthesis, and the DM and N translocation from their reserves at pre-anthesis. Post-anthesis assimilates accounted for 71–94% of the total grain yield among the barley varieties in 2014 and 53–81% in 2015. In contrast, the N reserved in vegetative tissues at anthesis contributed to barley grain N from 67% in the variety Union to 91% in the variety Marthe in 2014, and 71% in the variety Grace to 97% in the variety Shakira in 2015. The results concluded that photosynthetically derived assimilates at post-anthesis played an important role in determining grain size, whereas N reserves at pre-anthesis and N remobilization at post-anthesis probably determined the grain protein content of the malting barley. To achieve a high quality of malting barley grains in both grain size and protein content simultaneously, balancing photosynthetic assimilates at post-anthesis and N reserves at pre-anthesis and N remobilization should be considered as strategies for the combination of the selection of spring malting barley varieties together with agronomic N management.

Spectroscopic Estimation of N Concentration in Wheat Organs for Assessing N Remobilization Under Different Irrigation Regimes

Nitrogen (N) remobilization is a critical process that provides substantial N to winter wheat grains for improving yield productivity. Here, the remobilization of N from anthesis to maturity in two wheat cultivars under three irrigation regimes was measured and its relationship to organ N concentration was examined. Based on spectral data of organ powder samples, partial least squares regression (PLSR) models were calibrated to estimate N concentration (Nmass) and validated against laboratory-based measurements. Although spectral reflectance could accurately estimate Nmass, the PLSR-based Nmass-spectra predictive model was found to be organ-specific, organs at the top canopy (chaff and top three leaves) received the best predictions (R2 > 0.88). In addition, N remobilization efficiency (NRE) in the top two leaves and top third internode was highly correlated with its corresponding N concentration change (ΔNmass) with an R2 of 0.90. ΔNmass of the top first internode (TIN1) explained 78% variation of the whole-plant NRE. This study provides a proof of concept for estimating N concentration and assessing N remobilization using hyperspectral data of individual organs, which offers a non-chemical and low-cost approach to screen germplasms for an optimal NRE in drought-resistance breeding.

GARP transcription factors repress Arabidopsis nitrogen starvation response via ROS-dependent and -independent pathways

Plants need to cope with strong variations of nitrogen availability in the soil. Although many molecular players are being discovered concerning how plants perceive NO3− provision, it is less clear how plants recognize a lack of nitrogen. Following nitrogen removal, plants activate their nitrogen starvation response (NSR), which is characterized by the activation of very high-affinity nitrate transport systems (NRT2.4 and NRT2.5) and other sentinel genes involved in N remobilization such as GDH3. Using a combination of functional genomics via transcription factor perturbation and molecular physiology studies, we show that the transcription factors belonging to the HHO subfamily are important regulators of NSR through two potential mechanisms. First, HHOs directly repress the high-affinity nitrate transporters, NRT2.4 and NRT2.5. hho mutants display increased high-affinity nitrate transport activity, opening up promising perspectives for biotechnological applications. Second, we show that reactive oxygen species (ROS) are important to control NSR in wild-type plants and that HRS1 and HHO1 overexpressors and mutants are affected in their ROS content, defining a potential feed-forward branch of the signaling pathway. Taken together, our results define the relationships of two types of molecular players controlling the NSR, namely ROS and the HHO transcription factors. This work (i) up opens perspectives on a poorly understood nutrient-related signaling pathway and (ii) defines targets for molecular breeding of plants with enhanced NO3− uptake.

Ethephon Activates the Transcription of Senescence-Associated Genes and Nitrogen Mobilization in Grapevine Leaves (Vitis vinifera cv. Riesling)

Nitrogen (N) remobilization in the context of leaf senescence is of considerable importance for the viability of perennial plants. In late-ripening crops, such as Vitis vinifera, it may also affect berry ripening and fruit quality. Numerous studies on the model plant Arabidopsis thaliana have confirmed an involvement of the plant hormone ethylene in the regulation of senescence. However, ethylene research on grapevine was mostly focused on its involvement in berry ripening and stress tolerance until now. To investigate the effect of ethylene on the initiation, regulation, and progress of senescence-dependent N mobilization in grapevine leaves, we treated field-grown Vitis vinifera cv. Riesling vines with 25 mM ethephon at the end of berry ripening. Ethephon induced premature chlorophyll degradation and caused a shift of the leaf transcriptome equivalent to developmental leaf senescence. The upregulated metabolic processes covered the entire N remobilization process chain, altered the amino acid composition in the leaves, and resulted in an average 60% decrease in leaf N. Our findings increase the fundamental knowledge about the initiation and manipulation of leaf N remobilization in perennial woody plants by ethephon. This offers a methodological approach to the targeted induction of senescence and thus to an improvement in the N supply of grapes.

Physiological and Molecular Traits Associated with Nitrogen Uptake under Limited Nitrogen in Soft Red Winter Wheat

A sufficient nitrogen (N) supply is pivotal for high grain yield and desired grain protein content in wheat (Triticum aestivum L.). Elucidation of physiological and molecular mechanisms underlying nitrogen use efficiency (NUE) will enhance our ability to develop new N-saving varieties in wheat. In this study, we analyzed two soft red winter wheat genotypes, VA08MAS-369 and VA07W-415, with contrasting NUE under limited N. Our previous study demonstrated that higher NUE in VA08MAS-369 resulted from accelerated senescence and N remobilization in flag leaves at low N. The present study revealed that VA08MAS-369 also exhibited higher nitrogen uptake efficiency (NUpE) than VA07W-415 under limited N. VA08MAS-369 consistently maintained root growth parameters such as maximum root depth, total root diameter, total root surface area, and total root volume under N limitation, relative to VA07W-415. Our time-course N content analysis indicated that VA08MAS-369 absorbed N more abundantly than VA07W-415 after the anthesis stage at low N. More efficient N uptake in VA08MAS-369 was associated with the increased expression of genes encoding a two-component high-affinity nitrate transport system, including four NRT2s and three NAR2s, in roots at low N. Altogether, these results demonstrate that VA08MAS-369 can absorb N efficiently even under limited N due to maintained root development and increased function of N uptake. The ability of VA08MAS-369 in N remobilization and uptake suggests that this genotype could be a valuable genetic material for the improvement of NUE in soft red winter wheat.

Autophagy mediates grain yield and nitrogen stress resistance by modulating nitrogen remobilization in rice

Autophagy, a conserved cellular process in eukaryotes, has evolved to a sophisticated process to dispose of intracellular constituents and plays important roles in plant development, metabolism, and efficient nutrients remobilization under suboptimal nutrients conditions. Here, we show that OsATG8b, an AUTOPHAGY-RELATED8 (ATG8) gene in rice, was highly induced by nitrogen (N) starvation. Elevated expression of OsATG8b significantly increased ATG8 lipidation, autophagic flux, and grain yield in rice under both sufficient and deficient N conditions. Overexpressing of OsATG8b could greatly increase the activities of enzymes related to N metabolism. Intriguingly, the 15N-labeling assay further revealed that more N was remobilized to seeds in OsATG8b-overexpressing rice, which significantly increased the N remobilization efficiency (NRE), N harvest index, N utilization efficiency (NUE), and N uptake efficiency (NUpE). Conversely, the osatg8b knock-out mutants had the opposite results on these characters. The substantial transcriptional changes of the overexpressed transgenic lines indicated the presence of complex signaling to developmental, metabolic process, and hormone, etc. Excitingly, the transgenic rice under different backgrounds all similarly be boosted in yield and NUE with OsATG8b overexpression. This work provides an excellent candidate gene for improving N remobilization, utilization, and yield in crops simultaneously.

ACCELERATED CELL DEATH 6 Acts on Natural Leaf Senescence and Nitrogen Fluxes in Arabidopsis

As the last step of leaf development, senescence is a molecular process involving cell death mechanism. Leaf senescence is trigged by both internal age-dependent factors and environmental stresses. It must be tightly regulated for the plant to adopt a proper response to environmental variation and to allow the plant to recycle nutrients stored in senescing organs. However, little is known about factors that regulate both nutrients fluxes and plant senescence. Taking advantage of variation for natural leaf senescence between Arabidopsis thaliana accessions, Col-0 and Ct-1, we did a fine mapping of a quantitative trait loci for leaf senescence and identified ACCELERATED CELL DEATH 6 (ACD6) as the causal gene. Using two near-isogeneic lines, differing solely around the ACD6 locus, we showed that ACD6 regulates rosette growth, leaf chlorophyll content, as well as leaf nitrogen and carbon percentages. To unravel the role of ACD6 in N remobilization, the two isogenic lines and acd6 mutant were grown and labeled with 15N at the vegetative stage in order to determine 15N partitioning between plant organs at harvest. Results showed that N remobilization efficiency was significantly lower in all the genotypes with lower ACD6 activity irrespective of plant growth and productivity. Measurement of N uptake at vegetative and reproductive stages revealed that ACD6 did not modify N uptake efficiency but enhanced nitrogen translocation from root to silique. In this study, we have evidenced a new role of ACD6 in regulating both sequential and monocarpic senescences and disrupting the balance between N remobilization and N uptake that is required for a good seed filling.