Millet-inspired systems metabolic engineering of NUE in crops

The use of nitrogen (N) fertilizers in agriculture has a great ability to increase crop productivity. However, their excessive use has detrimental effects on the environment. Therefore, it is necessary to develop crop varieties with improved nitrogen use efficiency (NUE) that require less N but have substantial yields. Orphan crops such as millets are cultivated in limited regions and are well adapted to lower input conditions. Therefore, they serve as a rich source of beneficial traits that can be transferred into major crops to improve their NUE. This review highlights the tremendous potential of systems biology to unravel the enzymes and pathways involved in the N metabolism of millets, which can open new possibilities to generate transgenic crops with improved NUE.     

Cereal area and nitrogen use efficiency are drivers of future nitrogen fertilizer consumption

At a global scale, cereal yields and fertilizer N consumption have increased in a near-linear fashion during the past 40 years and are highly correlated with one another. However,large differences exist in historical trends of N fertilizer usage and nitrogen use efficiency (NUE)among regions, countries, and crops. The reasons for these differences must be understood to estimate future N fertilizer requirements. Global nitrogen needs will depend on: (i) changes in cropped cereal area and the associated yield increases required to meet increasing cereal demand from population and income growth, and (ii) changes in NUE at the farm level. Our analysis indicates that the anticipated 38% increase in global cereal demand by 2025 can be met by a 30% increase in N use on cereals, provided that the steady decline in cereal harvest area is halted and the yield response to applied N can be increased by 20%. If losses of cereal cropping area continue at the rate of the past 20 years (-0.33% per year) and NUE cannot be increased substantially, a 60% increase in global N use on cereals would be required to meet cereal demand. Interventions to increase NUE and reduce N losses to the environment must be accomplished at the farm- or field-scale through a combination of improved technologies and carefully crafted local policies that contribute to the adoption of improved N management; uniform regional or national directives are unlikey to be effective at both sustaining yield increases and improving NUE. Examples from several countries show that increases in NUE at rates of 1% per year or more can be achieved if adequate investments are made in research and extension. Failure to arrest the decrease in cereal crop area and to improve NUE in the world's most important agricultural systems will likely cause severe damage to environmental services at local, regional, and global scales due to a large increase in reactive N load in the environment.     

Cereal area and nitrogen use efficiency are drivers of future nitrogen fertilizer consumption

At a global scale, cereal yields and fertilizer N consumption have increased in a near-linear fashion during the past 40 years and are highly correlated with one another. However, large differences exist in historical trends of N fertilizer usage and nitrogen use efficiency (NUE) among regions, countries, and crops. The reasons for these differences must be understood to estimate future N fertilizer requirements. Global nitrogen needs will depend on: (i) changes in cropped cereal area and the associated yield increases required to meet increasing cereal demand from population and income growth, and (ii) changes in NUE at the farm level. Our analysis indicates that the anticipated 38% increase in global cereal demand by 2025 can be met by a 30% increase in N use on cereals, provided that the steady decline in cereal harvest area is halted and the yield response to applied N can be increased by 20%. If losses of cereal cropping area continue at the rate of the past 20 years (-0.33% per year) and NUE cannot be increased substantially, a 60% increase in global N use on cereals would be required to meet cereal demand. Interventions to increase NUE and reduce N losses to the environment must be accomplished at the farm- or field-scale through a combination of improved technologies and carefully crafted local policies that contribute to the adoption of improved N management; uniform regional or national directives are unlikey to be effective at both sustaining yield increases and improving NUE. Examples from several countries show that increases in NUE at rates of 1% per year or more can be achieved if adequate investments are made in research and extension. Failure to arrest the decrease in cereal crop area and to improve NUE in the world's most important agricultural systems will likely cause severe damage to environmental services at local, regional, and global scales due to a large increase in reactive N load in the environment.     

Cereal area and nitrogen use efficiency are drivers of future nitrogen fertilizer consumption

At a global scale, cereal yields and fertilizer N consumption have increased in a near-linear fashion during the past 40 years and are highly correlated with one another. However, large differences exist in historical trends of N fertilizer usage and nitrogen use efficiency (NUE) among regions, countries, and crops. The reasons for these differences must be understood to estimate future N fertilizer requirements. Global nitrogen needs will depend on: (i) changes in cropped cereal area and the associated yield increases required to meet increasing cereal demand from population and income growth, and (ii) changes in NUE at the farm level. Our analysis indicates that the anticipated 38% increase in global cereal demand by 2025 can be met by a 30% increase in N use on cereals, provided that the steady decline in cereal harvest area is halted and the yield response to applied N can be increased by 20%. If losses of cereal cropping area continue at the rate of the past 20 years (?0.33% per year) and NUE cannot be increased substantially, a 60% increase in global N use on cereals would be required to meet cereal demand. Interventions to increase NUE and reduce N losses to the environment must be accomplished at the farm-or field-scale through a combination of improved technologies and carefully crafted local policies that contribute to the adoption of improved N management; uniform regional or national directives are unlikey to be effective at both sustaining yield increases and improving NUE. Examples from several countries show that increases in NUE at rates of 1% per year or more can be achieved if adequate investments are made in research and extension. Failure to arrest the decrease in cereal crop area and to improve NUE in the world’s most important agricultural systems will likely cause severe damage to environmental services at local, regional, and global scales due to a large increase in reactive N load in the environment.     

Can less yield more? Is reducing nutrient input into the environment compatible with maintaining crop production?

Plant scientists have long recognized the need to develop crops that absorb and use nutrients more efficiently. Two approaches have been used to increase nutrient use efficiency (NUE) in crop plants. The first involves both traditional breeding and marker-assisted selection in an attempt to identify the genes involved. The second uses novel gene constructs designed to improve specific aspects of NUE. Here, we discuss some recent developments in the genetic manipulation of NUE in crop plants and argue that an improved understanding of the transition between nitrogen assimilation and nitrogen recycling will be important in applying this technology to increasing crop yields. Moreover, we emphasize the need to combine genetic and transgenic approaches to make significant improvements in NUE.     

The vegetative nitrogen response of sorghum lines containing different alleles for nitrate reductase and glutamate synthase

Improving the nitrogen (N) responsiveness of crops is crucial for food security and environmental sustainability, and breeding N use efficient (NUE) crops has to exploit genetic variation for this complex trait. We used reverse genetics to examine allelic variation in two N metabolism genes. In silico analysis of the genomes of 44 genetically diverse sorghum genotypes identified a nitrate reductase and a glutamate synthase gene that were under balancing selection in improved sorghum cultivars. We hypothesised that these genes are a potential source of differences in NUE, and selected parents and progeny of nested association mapping populations with different allelic combinations for these genes. Allelic variation was sourced from African (Macia) and Indian (ICSV754) genotypes that had been incorporated into the Australian elite parent R931945-2-2. Nine genotypes were grown for 30 days in a glasshouse and supplied with continuous limiting or replete N, or replete N for 27 days followed by 3 days of N starvation. Biomass, total N and nitrate contents were quantified together with gene expressions in leaves, stems and roots. Limiting N supply universally resulted in less shoot and root growth, increased root weight ratio and reduced tissue nitrate and total N concentrations. None of the tested genotypes exceeded growth or NUE of the elite parent R931945-2-2 indicating that the allelic combinations did not confer an advantage during early vegetative growth. Thus, the next steps for ascertaining potential effects on NUE include growing plants to maturity. We conclude that reverse genetics that take advantage of rapidly expanding genomic databases enable a systematic approach for developing N-efficient crops.     

Optimising non-destructive sampling methods to study nitrogen use efficiency throughout the growth-cycle of giant C4 crops

Plant and Soil - The improvement of nitrogen use efficiency (NUE) of crops allows crop nitrogen (N) demands to be met while reducing N supply, and so reducing excess N which has potential negative...     

Nitrogen use efficiency. 1. Uptake of nitrogen from the soil

The nitrogen use efficiency (NUE) of crop plants can be expressed very simply as the yield of nitrogen per unit of available nitrogen in the soil. This NUE can be divided into two processes: uptake efficiency, the ability of the plant to remove N from the soil normally present as nitrate or ammonium ions, and the utilisation efficiency, the ability of the plant to transfer the N to the grain, predominantly present as protein. In this article, we have highlighted the latest developments in the isolation and characterisation of the genes involved in the uptake of nitrogen from the soil.     

C3和C4植物的氮素利用机制

提高植物的氮素利用效率(NUE)不仅有利于保障全球粮食安全, 也是实现农业可持续发展的重要途径。近半个世纪以来, 植物氮素利用机理研究已取得重要进展, 但NUE的调控机制仍不明确, NUE的提高仍然十分有限。高等植物集光合碳素同化和氮素同化于一体, 只有碳氮代谢相互协调, 才能维持植物体内的碳氮平衡, 保证植物正常生长发育。由于C₃和C₄植物的光合氮素利用率(PNUE)存在差异, 对氮素的利用效率也会存在差异。为了更有效地提高作物的NUE, 须更全面地了解C₃和C₄植物对氮素吸收、转运、同化和信号转导等关键因子的功能和调控机制。此外, 面对大气CO₂浓度增高和全球气候变暖条件下的植物碳氮同化及其机理的研究也不容忽视。该文综述了C₃和C₄植物氮素利用关键因素的差异及其调控机制, 并对提高C₃禾本科作物氮素利用效率的遗传改良途径进行了展望。     

Nitrogen use efficiency. 2. Amino acid metabolism

In a previous article, we highlighted the latest developments in the isolation and characterisation of genes involved in the uptake of nitrogen from the soil, which might be used to improve the nitrogen use efficiency (NUE) of crop plants. In this article, we have concentrated on the genes controlling the enzymes of amino acid metabolism that may be involved in transferring nitrogen to the protein in the grain. Evidence is now accumulating from the use of knockout mutants, of the role of individual isoenzymes involved in amino acid metabolism, which are encoded by specific genes that are often members of a multigene family. In addition, a significant number of overexpressing plant lines have been obtained, which have increased activities of cytosol located, glutamine synthetase, asparagine synthetase and alanine aminotransferase that appear to have improved NUE.     

Cross-genome map based dissection of a nitrogen use efficiency ortho-metaQTL in bread wheat unravels concerted cereal genome evolution

Monitoring nitrogen use efficiency (NUE) in plants is becoming essential to maintain yield while reducing fertilizer usage. Optimized NUE application in major crops is essential for long-term sustainability of agriculture production. Here, we report the precise identification of 11 major chromosomal regions controlling NUE in wheat that co-localise with key developmental genes such as Ppd (photoperiod sensitivity), Vrn (vernalization requirement), Rht (reduced height) and can be considered as robust markers from a molecular breeding perspective. Physical mapping, sequencing, annotation and candidate gene validation of an NUE metaQTL on wheat chromosome 3B allowed us to propose that a glutamate synthase (GoGAT) gene that is conserved structurally and functionally at orthologous positions in rice, sorghum and maize genomes may contribute to NUE in wheat and other cereals. We propose an evolutionary model for the NUE locus in cereals from a common ancestral region, involving species specific shuffling events such as gene deletion, inversion, transposition and the invasion of repetitive elements.     

联合固氮菌的合成生物学研究进展

氮素是作物生长过程中最重要的元素,氮素缺乏将会严重影响作物生长。随着人类对粮食的需求量增加,化学氮肥的施用量越来越多。生物固氮在全球氮素循环中有着重要的作用,60%的氮来源于生物固氮。因此,生物固氮,尤其是能够在作物中定殖的联合固氮菌,最有可能代替氮肥成为粮食作物的主要氮源。长期以来,如何提高生物固氮效率以及在作物中实现生物固氮是生物学家的重要研究方向。合成生物学的出现和发展为能够生物固氮的研究带了新的机遇,有望缓解粮食作物对化学氮肥的大量需求。本文概述了固氮菌的种类、联合固氮菌中固氮基因岛的组成以及转录调控机理,阐述了合成生物学在生物固氮领域中的研究现状,对未来的联合固氮菌合成生物学的发展方向作出了展望。     

Plant nitrogen availability and crosstalk with phytohormones signallings and their biotechnology breeding application in crops

Nitrogen (N), one of the most important nutrients, limits plant growth and crop yields in sustainable agriculture system, in which phytohormones are known to play essential roles in N availability. Hence, it is not surprising that massive studies about the crosstalk between N and phytohormones have been constantly emerging. In this review, with the intellectual landscape of N and phytohormones crosstalk provided by the bibliometric analysis, we trace the research story of best-known crosstalk between N and various phytohormones over the last 20 years. Then, we discuss how N regulates various phytohormones biosynthesis and transport in plants. In reverse, we also summarize how phytohormones signallings modulate root system architecture (RSA) in response to N availability. Besides, we expand to outline how phytohormones signallings regulate uptake, transport, and assimilation of N in plants. Further, we conclude advanced biotechnology strategies, explain their application, and provide potential phytohormones-regulated N use efficiency (NUE) targets in crops. Collectively, this review provides not only a better understanding on the recent progress of crosstalk between N and phytohormones, but also targeted strategies for improvement of NUE to increase crop yields in future biotechnology breeding of crops.     

Mining of Candidate Maize Genes for Nitrogen Use Efficiency by Integrating Gene Expression and QTL Data

Breeding maize varieties for high nitrogen (N) use efficiency (NUE) by marker-assisted selection using NUE quantitative trait locus (QTL) or by genetic transfer of NUE-associated genes is a viable approach for increasing grain yield in N-limited production areas. In this investigation, we evaluated a set of introgression line populations under N supply and N deficiency conditions. From 42 QTLs for grain yield and yield components, 23 were identified under N supply conditions and 33 from N limited conditions. Meta-analysis of published maize NUE QTLs revealed 37 “consensus” QTLs, of which, 18 was detected under low N conditions. In addition, 258 unique ESTs associated with low N stress response, N uptake, transport, and assimilation were aligned on the maize genome by in silico mapping. Integrating the EST map with the QTL map has resulted in the identification of candidate NUE-associated genes of the following functional categories: N uptake, transport, and assimilation; carbon (C) metabolism and assimilation; and cascades of stress response and signal transduction genes. Nine candidates that have been introgressed into Ye478 significantly altered grain yield/yield components. It is suggested that the dynamics of interactions between C and N metabolism are important for maize yield. A high NUE variety should have a highly efficient C assimilation per unit N and actively express CO₂ assimilation-related genes under N-limited conditions.     

Association genetics of the parameters related to nitrogen use efficiency in Brassica juncea L.

Key message

Genome wide association studies allowed prediction of 17 candidate genes for association with nitrogen use efficiency. Novel information obtained may provide better understanding of genomic controls underlying germplasm variations for this trait in Indian mustard.

Abstract

Nitrogen use efficiency (NUE) of Indian mustard (Brassica juncea (L.) Czern & Coss.) is low and most breeding efforts to combine NUE with crop performance have not succeeded. Underlying genetics also remain unexplored. We tested 92 SNP-genotyped inbred lines for yield component traits, N uptake efficiency (NUPEFF), nitrogen utilization efficiency (NUTEFF), nitrogen harvest index (NHI) and NUE for two years at two nitrogen doses (N_o without added N and N₁₀₀ added @100 kg/ha). Genotypes IC-2489-88, M-633, MCP-632, HUJM 1080, GR-325 and DJ-65 recorded high NUE at low N. These also showed improved crop performance under high N. One determinate mustard genotype DJ-113 DT-3 revealed maximum NUTEFF. Genome wide association studies (GWAS) facilitated recognition of 17 quantitative trait loci (QTLs). Environment specificity was high. B-genome chromosomes (B02, B03, B05, B07 and B08) harbored many useful loci. We also used regional association mapping (RAM) to supplement results from GWAS. Annotation of the genomic regions around peak SNPs helped to predict several gene candidates for root architecture, N uptake, assimilation and remobilization. CAT9 (At1g05940) was consistently envisaged for both NUE and NUPEFF. Major N transporter genes, NRT1.8 and NRT3.1 were predicted for explaining variation for NUTEFF and NUPEFF, respectively. Most significant amino acid transporter gene, AAP1 appeared associated with NUE under limited N conditions. All these candidates were predicted in the regions of high linkage disequilibrium. Sequence information of the predicted candidate genes will permit development of molecular markers to aid breeding for high NUE.