Does energy cost constitute the primary cause of ammonium toxicity in plants?

Nitrate (NO3-) and ammonium (NH4+) are the main nitrogen (N) sources and key determinants for plant growth and development. In recent decades, NH4+, which is a double-sided N compound, has attracted considerable amounts of attention from researchers. Elucidating the mechanisms of NH4+ toxicity and exploring the means to overcome this toxicity are necessary to improve agricultural sustainability. In this review, we discuss the current knowledge concerning the energy consumption and production underlying NH4+ metabolism and toxicity in plants, such as N uptake; assimilation; cellular pH homeostasis; and functions of the plasma membrane (PM), vacuolar H+-ATPase and H+-pyrophosphatase (H+-PPase). We also discuss whether the overconsumption of energy is the primary cause of NH4+ toxicity or constitutes a fundamental strategy for plants to adapt to high-NH4+ stress. In addition, the effects of regulators on energy production and consumption and other physiological processes are listed for evaluating the possibility of high energy costs associated with NH4+ toxicity. This review is helpful for exploring the tolerance mechanisms and for developing NH4+-tolerant varieties as well as agronomic techniques to alleviate the effects of NH4+ stress in the field.

Nitrate alleviates ammonium toxicity in wheat (Triticum aestivum L.) by regulating tricarboxylic acid cycle and reducing rhizospheric acidification and oxidative damage.

Ammonium (NH4+) is one of the most important nutrients required by plants. However, a high concentration of NH4+ as the sole nitrogen source suppresses plant growth. Although nitrate (NO3-) can alleviate NH4+ toxicity, the mechanisms underlying this ability have not been fully elucidated. In this study, wheat plants were cultivated in hydroponic solution with 7.5 mM NO3- (control), 7.5 mM NH4+ (sole ammonium, SA) or 7.5 mM NH4+ plus 1.0 mM NO3- (ammonium and nitrate, AN). The results showed that compared with the control, the SA treatment significantly decreased root growth, protein content and the concentrations of most intermediates and the activity of enzymes from the tricarboxylic acid (TCA) cycle. Moreover, increased the activity of plasma membrane H+-ATPase and the rate of H+ efflux along roots, caused solution acidification, and increased the activity of mitochondrial respiratory chain complexes I-IV and the contents of protein-bound carbonyls and malondialdehyde in roots. SA treatment induced ultrastructure disruption and reduced the viability of root cells. Compared with the SA treatment, the AN treatment increased root growth, protein content, the concentrations of most intermediates and the activity of enzymes from the TCA cycle. Furthermore, AN treatment decreased the rate of H+ efflux, retarded medium acidification, decreased protein carbonylation and lipid peroxidation in roots and relieved ultrastructure disruption and increased the viability of root cells. Taken together, these results indicate that NO3--dependent alleviation of NH4+ toxicity in wheat seedlings is closely associated with physiological processes that mediate TCA cycle, relieve rhizospheric acidification and decrease the production of ROS and oxidative damage.

Genome-wide characterization of the abscisic acid-, stress- and ripening-induced (ASR) gene family in wheat (Triticum aestivum L.).

BACKGROUND: Abscisic acid-, stress-, and ripening-induced (ASR) genes are a class of plant specific transcription factors (TFs), which play important roles in plant development, growth and abiotic stress responses. The wheat ASRs have not been described in genome-wide yet. METHODS: We predicted the transmembrane regions and subcellular localization using the TMHMM server, and Plant-mPLoc server and CELLO v2.5, respectively. Then the phylogeny tree was built by MEGA7. The exon-intron structures, conserved motifs and TFs binding sites were analyzed by GSDS, MEME program and PlantRegMap, respectively. RESULTS: In wheat, 33ASR genes were identified through a genome-wide survey and classified into six groups. Phylogenetic analyses revealed that the TaASR proteins in the same group tightly clustered together, compared with those from other species. Duplication analysis indicated that the TaASR gene family has expanded mainly through tandem and segmental duplication events. Similar gene structures and conserved protein motifs of TaASRs in wheat were identified in the same groups. ASR genes contained various TF binding cites associated with the stress responses in the promoter region. Gene expression was generally associated with the expected group-specific expression pattern in five tissues, including grain, leaf, root, spike and stem, indicating the broad conservation of ASR genes function during wheat evolution. The qRT-PCR analysis revealed that several ASRs were up-regulated in response to NaCl and PEG stress. CONCLUSION: We identified ASR genes in wheat and found that gene duplication events are the main driving force for ASR gene evolution in wheat. The expression of wheat ASR genes was modulated in responses to multiple abiotic stresses, including drought/osmotic and salt stress. The results provided important information for further identifications of the functions of wheat ASR genes and candidate genes for high abiotic stress tolerant wheat breeding.

Genome-wide characterization of the abscisic acid-, stress- and ripening-induced (ASR) gene family in wheat (Triticum aestivum L)

BACKGROUND: Abscisic acid-, stress-, and ripening-induced (ASR) genes are a class of plant specific transcription factors (TFs), which play important roles in plant development, growth and abiotic stress responses. The wheat ASRs have not been described in genome-wide yet. METHODS: We predicted the transmembrane regions and subcellular localization using the TMHMM server, and Plant-mPLoc server and CELLO v2.5, respectively. Then the phylogeny tree was built by MEGA7. The exon-intron structures, conserved motifs and TFs binding sites were analyzed by GSDS, MEME program and PlantRegMap, respectively. RESULTS: In wheat, 33ASR genes were identified through a genome-wide survey and classified into six groups. Phylogenetic analyses revealed that the TaASR proteins in the same group tightly clustered together, compared with those from other species. Duplication analysis indicated that the TaASR gene family has expanded mainly through tandem and segmental duplication events. Similar gene structures and conserved protein motifs of TaASRs in wheat were identified in the same groups. ASR genes contained various TF binding cites associated with the stress responses in the promoter region. Gene expression was generally associated with the expected group-specific expression pattern in five tissues, including grain, leaf, root, spike and stem, indicating the broad conservation of ASR genes function during wheat evolution. The qRT-PCR analysis revealed that several ASRs were up-regulated in response to NaCl and PEG stress. CONCLUSION: We identified ASR genes in wheat and found that gene duplication events are the main driving force for ASR gene evolution in wheat. The expression of wheat ASR genes was modulated in responses to multiple abiotic stresses, including drought/osmotic and salt stress. The results provided important information for further identifications of the functions of wheat ASR genes and candidate genes for high abiotic stress tolerant wheat breeding.

Excessive nitrogen application dampens antioxidant capacity and grain filling in wheat as revealed by metabolic and physiological analyses.

In this study, field-grown wheat (Triticum aestivum L.) was treated with normal (Nn) and excessive (Ne) levels of fertilizer N. Results showed that Ne depressed the activity of superoxide dismutase and peroxidase and increased the accumulation of reactive oxygen species (ROS) and malondialdehyde. The normalized difference vegetation index (NDVI) was higher under Ne at anthesis and medium milk but similar at the early dough stage and significantly lower at the hard dough stage than that under Nn. The metabolomics analysis of the leaf responses to Ne during grain filling showed 99 metabolites that were different between Ne and Nn treatments, including phenolic and flavonoid compounds, amino acids, organic acids and lipids, which are primarily involved in ROS scavenging, N metabolism, heat stress adaptation and disease resistance. Organic carbon (C) and total N contents were affected by the Ne treatment, with lower C/N ratios developing after medium milk. Ultimately, grain yields decreased with Ne. Based on these data, compared with the normal N fertilizer treatment, we concluded that excessive N application decreased the ability to scavenge ROS, increased lipid peroxidation and caused significant metabolic changes disturbing N metabolism, secondary metabolism and lipid metabolism, which led to reduced grain filling in wheat.

Effects of high NH(+) 4 on K(+) uptake, culm mechanical strength and grain filling in wheat.

It is well established that a high external NH(+) 4 concentration depresses many processes in plant development, but the underlying mechanisms are still not well understood. To determine whether the negative effects of high levels of NH(+) 4 are related to competitive cation uptake, wheat was grown in a field with moderate (18 g N m(-2)) and high (30 g N m(-2)) supplies of NH(+) 4 in the presence or absence of additional K(+) (6 g K2O m(-2)) to examine culm mechanical strength, the main components of the vascular bundle, nitrogen (N) remobilization and the grain-filling rate. The results indicated that an excessive supply of NH(+) 4 significantly decreased culm mechanical strength, the cellulose and lignin contents of vascular bundles, the N remobilization efficiency (NRE) and the grain-filling rate compared with a moderate level of NH(+) 4. The additional provision of K(+) considerably alleviated these negative effects of high NH(+) 4, resulting in a 19.41-26.95% increase in culm mechanical strength during grain filling and a 34.59% increase in the NRE. An assay using the scanning ion-selective electrode technique (SIET) showed that the net rate of transmembrane K(+) influx decreased by 84.62%, and measurements using flame photometry demonstrated that the K(+) content decreased by 36.13% in wheat plants subjected to high NH(+) 4. This study indicates that the effects of high NH(+) 4 on culm mechanical strength, cellulose and lignin contents, the NRE and the grain-filling rate are probably associated with inhibition of K(+) uptake in wheat.

The structural and photosynthetic characteristics of the exposed peduncle of wheat (Triticum aestivum L.): an important photosynthate source for grain-filling.

BACKGROUND: In wheat (Triticum aestivum L), the flag leaf has been thought of as the main source of assimilates for grain growth, whereas the peduncle has commonly been thought of as a transporting organ. The photosynthetic characteristics of the exposed peduncle have therefore been neglected. In this study, we investigated the anatomical traits of the exposed peduncle during wheat grain ontogenesis, and we compared the exposed peduncle to the flag leaf with respect to chloroplast ultrastructure, photosystem II (PSII) quantum yield, and phosphoenolpyruvate carboxylase (PEPCase; EC 4.1.1.31) activity. RESULTS: Transmission electron microscope observations showed well-developed chloroplasts with numerous granum stacks at grain-filling stages 1, 2 and 3 in both the flag leaf and the exposed peduncle. In the exposed peduncle, the membranes constituting the thylakoids were very distinct and plentiful, but in the flag leaf, there was a sharp breakdown at stage 4 and complete disintegration of the thylakoid membranes at stage 5. PSII quantum yield assays revealed that the photosynthetic efficiency remained constant at stages 1, 2 and 3 and then declined in both organs. However, the decline occurred more dramatically in the flag leaf than in the exposed peduncle. An enzyme assay showed that at stages 1 and 2 the PEPCase activity was lower in the exposed peduncle than in the flag leaf; but at stages 3, 4 and 5 the value was higher in the exposed peduncle, with a particularly significant difference observed at stage 5. Subjecting the exposed part of the peduncle to darkness following anthesis reduced the rate of grain growth. CONCLUSION: Our results suggest that the exposed peduncle is a photosynthetically active organ that produces photosynthates and thereby makes a crucial contribution to grain growth, particularly during the late stages of grain-filling.