Engineering nitrogen use efficient crop plants: the current status

In the last 40 years the amount of synthetic nitrogen (N) applied to crops has risen drastically, resulting in significant increases in yield but with considerable impacts on the environment. A requirement for crops that require decreased N fertilizer levels has been recognized in the call for a ‘Second Green Revolution’ and research in the field of nitrogen use efficiency (NUE) has continued to grow. This has prompted a search to identify genes that improve the NUE of crop plants, with candidate NUE genes existing in pathways relating to N uptake, assimilation, amino acid biosynthesis, C/N storage and metabolism, signalling and regulation of N metabolism and translocation, remobilization and senescence. Herein is a review of the approaches taken to determine possible NUE candidate genes, an overview of experimental study of these genes as effectors of NUE in both cereal and non‐cereal plants and the processes of commercialization of enhanced NUE crop plants. Patents issued regarding increased NUE in plants as well as gene pyramiding studies are also discussed as well as future directions of NUE research.     

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.     

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。     

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.     

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.

     

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.     

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.     

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...     

The challenge of improving nitrogen use efficiency in crop plants: towards a more central role for genetic variability and quantitative genetics within integrated approaches

In this review, recent developments and future prospects of obtaining a better understanding of the regulation of nitrogen use efficiency in the main crop species cultivated in the world are presented. In these crops, an increased knowledge of the regulatory mechanisms controlling plant nitrogen economy is vital for improving nitrogen use efficiency and for reducing excessive input of fertilizers, while maintaining an acceptable yield. Using plants grown under agronomic conditions at low and high nitrogen fertilization regimes, it is now possible to develop whole-plant physiological studies combined with gene, protein, and metabolite profiling to build up a comprehensive picture depicting the different steps of nitrogen uptake, assimilation, and recycling to the final deposition in the seed. A critical overview is provided on how understanding of the physiological and molecular controls of N assimilation under varying environmental conditions in crops has been improved through the use of combined approaches, mainly based on whole-plant physiology, quantitative genetics, and forward and reverse genetics approaches. Current knowledge and prospects for future agronomic development and application for breeding crops adapted to lower fertilizer input are explored, taking into account the world economic and environmental constraints in the next century.     

Can genetic manipulation of plant nitrogen assimilation enzymes result in increased crop yield and greater N-use efficiency? An assessment

The literature on the relations between plant nitrogen (N) assimilation enzymes and plant/crop N assimilation, growth and yield is reviewed to assess if genetic manipulation of the activities of N assimilation enzymes can result in increased yield and/or increased N use efficiency. The available data indicate that (I) levels of N assimilation enzymes do not limit primary N assimilation and hence yield; (II) root or shoot nitrate assimilation can have advantages under specific environmental conditions; (III) for cereals, cytosolic glutamine synthetase (GS1) is a key enzyme in the mobilisation of N from senescing leaves and its activity in senescing leaves is positively related to yield; and (TV) for rice (Oryza sativd), NADH-glutamate synthase (NADH-GOGAT) is important in the utilisation of N in grain filling and its activity in developing grains is positively related to yield. In our opinion, selection of plants, from either a genetically manipulated population or genetic resources, with expression of nitrate reductase/nitrite reductase primarily in the root or shoot should increase plant/crop growth and hence yield under specific environmental conditions. In addition for cereals the selection of plants with high GS1 in senescing leaves and in some cases high NADH-GOGAT in developing grains could help maximise the retrieval of plant N in seeds.     

Agronomic improvements through the genetic and physiological regulation of nitrogen uptake in wheat (Triticum aestivum L.)

Nitrogen (N) uptake is the first step in nitrate assimilation, and efficient N uptake is essential for plant growth, especially for protein biosynthesis and photosynthetic activities. In cereals, improved N uptake is closely coupled with an increase in nitrogen use efficiency (NUE) and yield improvements. Because wheat (Triticum aestivum L.) is a leading crop worldwide, a better understanding of N uptake regulation in wheat is vital to improving NUE and developing sustainable agricultural systems. However, detailed information regarding the biological mechanisms that are responsible for the more efficient uptake of ambient N by wheat is limited. This review presents recent developments in the biological mechanisms of N uptake in wheat, including plant growth regulations, fundamental roles of root systems, interactions between N species, and genetic controls. Specifically, this paper provides a number of potential strategies that can be used to increase wheat N uptake. The information provided here may guide N fertilizer management during wheat production and further elucidate the plant regulatory mechanisms that are involved in N uptake, which can thereby increase wheat NUE.     

Identification of quantitative trait loci for nitrogen use efficiency in maize

Intensively managed crop systems are normally dependent on nitrogen input to maximize yield potential. Improvements in nitrogen- use efficiency (NUE) in crop plants may support the development of cropping systems that are more economically efficient and environment friendly. The objective of this study was to map and characterize quantitative trait loci (QTL) for NUE in a maize population. In preliminary experiments, inbred lines contrasting for NUE were identified and were used to generate populations of F2:3 families for genetic study. A total of 214 F2:3 families were evaluated in replicated trials under high nitrogen (280 kg/ha) and low nitrogen (30 kg/ha) conditions in 1996 and 1997. Analysis of ear-leaf area, plant height, grain yield, ears per plant, kernels number per ear, and kernel weight indicated significant genetic variation among F2:3 families. The heritability of these traits was found to be high (h2=0.57–0.81). The mapping population were genotyped using a set of 99 restriction fragment length polymorphism (RFLP) markers. A linkage map of these markers was developed and used to identify QTL. Between two and six loci were found to be associated with each trait. The correspondence of several genomic regions with traits measured under nitrogen limited conditions suggests the presence of QTL associated with NUE. QTLs will help breeders to improve their maize ideotype of a low-nitrogen efficiency by identifying those constitutive and adaptive traits involved in the expression of traits significantly correlated with yield, such as ear leaf area and number of ears per plant. This revised version was published online in June 2006 with corrections to the Cover Date.     

Nitrogen use efficiencies of spring barley grown under varying nitrogen conditions in the field and growth chamber

Background and Aims

Nitrogen-use efficiency (NUE) of cereals needs to be improved by nitrogen (N) management, traditional plant breeding methods and/or biotechnology, while maintaining or, optimally, increasing crop yields. The aims of this study were to compare spring-barley genotypes grown on different nitrogen levels in field and growth-chamber conditions to determine the effects on N uptake (NUpE) and N utilization efficiency (NUtE) and ultimately, NUE.

Methods

Morphological characteristics, seed yield and metabolite levels of 12 spring barley (Hordeum vulgare) genotypes were compared when grown at high and low nitrogen levels in field conditions during the 2007 and 2008 Canadian growing seasons, and in potted and hydroponic growth-chamber conditions. Genotypic NUpE, NUtE and NUE were calculated and compared between field and growth-chamber environments.

Key Results

Growth chamber and field tests generally showed consistent NUE characteristics. In the field, Vivar, Excel and Ponoka, showed high NUE phenotypes across years and N levels. Vivar also had high NUE in growth-chamber trials, showing NUE across complex to simplistic growth environments. With the high NUE genotypes grown at low N in the field, NUtE predominates over NUpE. N metabolism-associated amino acid levels were different between roots (elevated glutamine) and shoots (elevated glutamate and alanine) of hydroponically grown genotypes. In field trials, metabolite levels were different between Kasota grown at high N (elevated glutamine) and Kasota at low N plus Vivar at either N condition.

Conclusions

Determining which trait(s) or gene(s) to target to improve barley NUE is important and can be facilitated using simplified growth approaches to help determine the NUE phenotype of various genotypes. The genotypes studied showed similar growth and NUE characteristics across field and growth-chamber tests demonstrating that simplified, low-variable growth environments can help pinpoint genetic targets for improving spring barley NUE.     

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.     

Growth and nitrogen use in Xanthium canadense grown in an open or in a dense stand

Plants develop branches profusely when grown solitarily, while less so when grown in a dense stand. Such changes in architecture are associated with changes in dry mass allocation and nitrogen use. Here, we studied what traits in plant growth and nitrogen use were influenced by different light climates in the stand. Annual plants (Xanthium canadense) were grown solitarily or in a dense stand. Dry mass growth was analyzed as the product of the net assimilation rate (NAR) and leaf area (LA). Nitrogen use efficiency (NUE) was analyzed as the product of nitrogen productivity (NP) and the mean residence time (MRT) of nitrogen. These growth variables were further factorized into their components. Solitary plants maintained a high NAR, whereas plants in the dense stand decreased the NAR due to mutual shading. Plants in the dense stand developed a larger LA with a higher specific leaf area than solitary plants. Solitary plants had higher NUE due to higher NP. A temporal increase in NUE was attributed to the increase in MRT of nitrogen. Light climate was different between solitary and dense-stand plants, but they took up a comparable amount of nitrogen and used it differently in response to the given light climate. NUE was thus demonstrated to be a useful tool for analyzing the mechanism leading to different N use in plant growth.     

A multi-environmental study of recent breeding progress on nitrogen use efficiency in wheat (Triticum aestivum L.)

Key message

By comparing 195 varieties in eight trials, this study assesses nitrogen use efficiency improvement in high and low nitrogen conditions in European winter wheat over the last 25 years.

Abstract

In a context where European agriculture practices have to deal with environmental concerns and nitrogen (N) fertiliser cost, nitrogen use efficiency (NUE) has to be improved. This study assessed genetic progress in winter wheat (Triticum aestivum L.) NUE. Two hundred and twenty-five European elite varieties were tested in four environments under two levels of N. Global genetic progress was assessed on additive genetic values and on genotype × N interaction, covering 25 years of European breeding. To avoid sampling bias, quality, precocity and plant height were added as covariates in the analyses when needed. Genotype × environment interactions were highly significant for all the traits studied to such an extent that no additive genetic effect was detected on N uptake. Genotype × N interactions were significant for yield, grain protein content (GPC), N concentration in straw, N utilisation, and NUE. Grain yield improvement (+0.45 % year^?1) was independent of the N treatment. GPC was stable, thus grain nitrogen yield was improved (+0.39 % year^?1). Genetic progress on N harvest index (+0.12 % year^?1) and on N concentration in straw (?0.52 % year^?1) possibly revealed improvement in N remobilisation. There has been an improvement of NUE additive genetic value (+0.33 % year^?1) linked to better N utilisation (+0.20 % year^?1). Improved yield stability was detected as a significant improvement of NUE in low compared to high N conditions. The application of these results to breeding programs is discussed.