Low Nitrogen Stress Research Articles

OsCIPK2 mediated rice root microorganisms and metabolites to improve plant nitrogen uptake

Crop roots are colonized by large numbers of microorganisms, collectively known as the root-microbiome, which modulate plant growth, development and contribute to elemental nutrient uptake. In conditions of nitrogen limitation, the over-expressed Calcineurin B-like interacting protein kinase 2 (OsCIPK2) gene with root-specific promoter (RC) has been shown to enhance growth and nitrogen uptake in rice. Analysis of root-associated bacteria through high-throughput sequencing revealed that OsCIPK2 has a significant impact on the diversity of the root microbial community under low nitrogen stress. The quantification of nifH gene expression demonstrated a significant enhancement in nitrogen-fixing capabilities in the roots of RC transgenetic rice. Synthetic microbial communities (SynCom) consisting of six nitrogen-fixing bacterial strains were observed to be enriched in the roots of RC, leading to a substantial improvement in rice growth and nitrogen uptake in nitrogen-deficient soils. Forty and twenty-three metabolites exhibiting differential abundance were identified in the roots and rhizosphere soils of RC transgenic rice compared to wild-type (WT) rice. These findings suggest that OSCIPK2 plays a role in restructuring the microbial community in the roots through the regulation of metabolite synthesis and secretion. Further experiments involving the exogenous addition of citric acid revealed that an optimal concentration of this compound facilitated the growth of nitrogen-fixing bacteria and substantially augmented their population in the soil, highlighting the importance of citric acid in promoting nitrogen fixation under conditions of low nitrogen availability. These findings suggest that OsCIPK2 plays a role in enhancing nitrogen uptake by rice plants from the soil by influencing the assembly of root microbial communities, thereby offering valuable insights for enhancing nitrogen utilization in rice cultivation.

Overexpression of oilseed rape trehalose-6-phosphate synthesis gene BnaC02.TPS8 confers sensitivity to low nitrogen and high sucrose-induced anthocyanin accumulation in Arabidopsis.

Overexpression of BnaC02.TPS8 increased low N and high sucrose-induced anthocyanin accumulation. Anthocyanin plays a crucial role in safeguarding photosynthetic tissues against high light, UV radiation, and oxidative stress. Their accumulation is triggered by low nitrogen (N) stress and elevated sucrose levels in Arabidopsis. Trehalose-6-phosphate (T6P) serves as a pivotal signaling molecule, sensing sucrose availability, and carbon (C) metabolism. However, the mechanisms governing the regulation of T6P synthase (TPS) genes responsible for anthocyanin accumulation under conditions of low N and high sucrose remain elusive. In a previous study, we demonstrated the positive impact of a cytoplasm-localized class II TPS protein 'BnaC02.TPS8' on photosynthesis and seed yield improvement in Brassica napus. The present research delves into the biological role of BnaC02.TPS8 in response to low N and high sucrose. Ectopic overexpression of BnaC02.TPS8 in Arabidopsis seedlings resulted in elevated shoot T6P levels under N-sufficient conditions, as well as an increased carbon-to-nitrogen (C/N) ratio, sucrose accumulation, and starch storage under low N conditions. Overexpression of BnaC02.TPS8 in Arabidopsis heightened sensitivity to low N stress and high sucrose levels, accompanied by increased anthocyanin accumulation and upregulation of genes involved in flavonoid biosynthesis and regulation. Metabolic profiling revealed increased levels of intermediate products of carbon metabolism, as well as anthocyanin and flavonoid derivatives in BnaC02.TPS8-overexpressing Arabidopsis plants under low N conditions. Furthermore, yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) analyses demonstrated that BnaC02.TPS8 interacts with both BnaC08.TPS9 and BnaA01.TPS10. These findings contribute to our understanding of how TPS8-mediated anthocyanin accumulation is modulated under low N and high sucrose conditions.

Genome-wide characterization of TCP family and their potential roles in abiotic stress resistance of oat (Avena sativa L.).

The TCP gene family members play multiple functions in plant growth and development and were named after the first three family members found in this family, TB1 (TEOSINTE BRANCHED 1), CYCLOIDEA (CYC), and Proliferating Cell Factor 1/2 (PCF1/2). Nitrogen (N) is a crucial element for forage yield; however, over-application of N fertilizer can increase agricultural production costs and environmental stress. Therefore, the discovery of low N tolerance genes is essential for the genetic improvement of superior oat germplasm and ecological protection. Oat (Avena sativa L.), is one of the world's staple grass forages, but no genome-wide analysis of TCP genes and their roles in low-nitrogen stress has been performed. This study identified the oat TCP gene family members using bioinformatics techniques. It analyzed their phylogeny, gene structure analysis, and expression patterns. The results showed that the AsTCP gene family includes 49 members, and most of the AsTCP-encoded proteins are neutral or acidic proteins; the phylogenetic tree classified the AsTCP gene family members into three subfamilies, and each subfamily has different conserved structural domains and functions. In addition, multiple cis-acting elements were detected in the promoter of the AsTCP genes, which were associated with abiotic stress, light response, and hormone response. The 49 AsTCP genes identified from oat were unevenly distributed on 18 oat chromosomes. The results of real-time quantitative polymerase chain reaction (qRT-PCR) showed that the AsTCP genes had different expression levels in various tissues under low nitrogen stress, which indicated that these genes (such as AsTCP01, AsTCP03, AsTCP22, and AsTCP38) played multiple roles in the growth and development of oat. In conclusion, this study analyzed the AsTCP gene family and their potential functions in low nitrogen stress at the genome-wide level, which lays a foundation for further analysis of the functions of AsTCP genes in oat and provides a theoretical basis for the exploration of excellent stress tolerance genes in oat. This study provides an essential basis for future in-depth studies of the TCP gene family in other oat genera and reveals new research ideas to improve gene utilization.

Physiological and Transcriptional Characteristics of Banana Seedlings in Response to Nitrogen Deficiency Stress

Nitrogen is a crucial element for the growth and development of plants, directly affecting crop growth and yield. To investigate the physiological and molecular mechanism of nitrogen-deficiency stress, we conducted an investigation into the effects of different nitrogen levels on the growth, photosynthetic characteristics, and gene transcription levels of banana seedlings. Compared with the control group with normal nitrogen levels (NN), the height of plants receiving Reduced-N (NR), Low-N (LN), and N-Free (NF) treatments was decreased by 0.45 cm, 2.5 cm, and 3.25 cm, respectively. Their dry weight was reduced by 1.63 g, 2.99 g, and 2.88 g, respectively. Conversely, the dry weight of the underground plant part in the LN and NF treatment groups exhibited an increase of 0.13 g and 0.16 g, respectively. Regarding photosynthetic characteristics, the Specialty Products Agricultural Division (SPAD) values of the NR, LN, and NF treatments showed reductions of 15.5%, 30.4%, and 35.9%, respectively, compared with those of the control treatments. The values of maximum photosynthetic efficiency (Fv/Fm), actual photosynthetic efficiency (Y(Ⅱ)), and relative electron transfer (ETR) of the banana seedlings decreased to different degrees after NR, LN, and NF treatment, and their values were positively correlated with N levels. Gene transcription analysis showed that N transport-related proteins, including NRT1.7, NRT2.3a, NRT2.3b, and NRT2.5, were significantly up-regulated to increase the nitrogen absorption capacity of plant roots. On the other hand, various transcription factors including GRAS, MYB, and WRKY were notably up-regulated, facilitating root growth and the expanding root absorption area, thereby enhancing nitrogen uptake. Furthermore, genes associated with endogenous hormone metabolic pathways such as gibberellin (GA), strigolactone (SL), and brassinosteroids (BR) were activated in banana plants subjected to low nitrogen stress, enhancing the plant’s ability to adapt to nitrogen-deficient conditions. These findings offer valuable insights into understanding the transcriptional regulatory mechanisms governing banana responses to low nitrogen stress and breeding new varieties with improved nutrient utilization.

MiR164-MhNAC1 regulates apple root nitrogen uptake under low nitrogen stress.

Nitrogen is an essential nutrient for plant growth and serves as a signaling molecule to regulate gene expression inducing physiological, growth and developmental responses. An excess or deficiency of nitrogen may have adverse effects on plants. Studying nitrogen uptake will help us understand the molecular mechanisms of utilization for targeted molecular breeding. Here, we identified and functionally validated an NAC (NAM-ATAF1/2-CUC2) transcription factor based on the transcriptomes of two apple rootstocks with different nitrogen uptake efficiency. NAC1, a target gene of miR164, directly regulates the expression of the high-affinity nitrate transporter (MhNRT2.4) and citric acid transporter (MhMATE), affecting root nitrogen uptake. To examine the role of MhNAC1 in nitrogen uptake, we produced transgenic lines that overexpressed or silenced MhNAC1. Silencing MhNAC1 promoted nitrogen uptake and citric acid secretion in roots, and enhanced plant tolerance to low nitrogen conditions, while overexpression of MhNAC1 or silencing miR164 had the opposite effect. This study not only revealed the role of the miR164-MhNAC1 module in nitrogen uptake in apple rootstocks but also confirmed that citric acid secretion in roots affected nitrogen uptake, which provides a research basis for efficient nitrogen utilization and molecular breeding in apple.

Fulvic acid alleviates the stress of low nitrogen on maize by promoting root development and nitrogen metabolism.

The potential of fulvic acid (FA) to improve plant growth has been acknowledged, but its effect on plant growth and nutrient uptake under nutrient stress remains unclear. This study investigated the effects of different FA application rates on maize growth and nitrogen utilization under low nitrogen stress. The results showed that under low nitrogen stress, FA significantly stimulated maize growth, particularly root development, biomass, and nitrogen content. The enhanced activity levels of key enzymes in nitrogen metabolism were observed, along with differential gene expression in maize, which enriched nitrogen metabolism, amino acid metabolism and plant hormone metabolism. The application of FA regulated the hormones' level, reduced abscisic acid content in leaves and Me-JA content in roots, and increased auxin and zeatin ribose content in leaves. This study concludes that, by promoting root development, nitrogen metabolism, and hormone metabolism, an appropriate concentration of FA can enhance plant tolerance to low nitrogen conditions and improve nitrogen use efficiency.

Brassinosteroids improve the redox state of wheat florets under low-nitrogen stress and alleviate degeneration

Reducing nitrogen application rates can mitigate issues such as environmental degradation and resource wastage. However, it can also exacerbate problems such as wheat floret degeneration, leading to reduced yields. Therefore, investigating wheat floret degeneration mechanisms under low nitrogen stress and identifying mitigation measures are conducive to achieving high yields and sustainable development. To investigate the physiological mechanism of low nitrogen stress affecting wheat floret degradation and whether exogenous brassinosteroids can alleviate this stress, three nitrogen application rates (N0, no nitrogen application; N1, 120 kg ha-1 pure nitrogen; and N2, 240 kg ha-1 pure nitrogen) and exogenous spraying experiments (N0CK, no nitrogen with water spraying; N0BR, no nitrogen with 24-epibrassinolide (an active brassinosteroids) spraying; and N1, 120 kg ha-1 pure nitrogen with water spraying) were designed. The results indicated that low nitrogen stress induced a large amount of reactive oxygen species generation. Although wheat spikes synthesized flavonoids to combat oxidative stress, their energy metabolism (glycolysis and tricarboxylic acid cycle) and ascorbate-glutathione cycle were inhibited, keeping reactive oxygen levels elevated within the spike, inducing cell death and exacerbating floret degeneration. Furthermore, brassinosteroids played a role in regulating wheat floret degeneration under low-nitrogen stress. Exogenous foliar spraying of 24-epibrassinolide promoted energy metabolism and the ascorbate-glutathione cycle within the spike, enhancing energy charge and effectively mitigating a portion of reactive oxygen induced by low nitrogen stress, thereby alleviating floret degeneration caused by low nitrogen stress. In summary, low-nitrogen stress disrupts the redox homeostasis of wheat spikes, leading to floret degeneration. Brassinosteroids alleviate floret degeneration by improving the redox state of wheat spikes. This research provides theoretical support for balancing the contradiction between high yields and sustainable development and is beneficial for the application of low nitrogen in production.

Transcriptome Profiling Reveals the Gene Network Responding to Low Nitrogen Stress in Wheat

As one of the essential nutrients for plants, nitrogen (N) has a major impact on the yield and quality of wheat worldwide. Due to chemical fertilizer pollution, it has become increasingly important to improve crop yield by increasing N use efficiency (NUE). Therefore, understanding the response mechanisms to low N (LN) stress is essential for the regulation of NUE in wheat. In this study, LN stress significantly accelerated wheat root growth, but inhibited shoot growth. Further transcriptome analysis showed that 8468 differentially expressed genes (DEGs) responded to LN stress. The roots and shoots displayed opposite response patterns, of which the majority of DEGs in roots were up-regulated (66.15%; 2955/4467), but the majority of DEGs in shoots were down-regulated (71.62%; 3274/4565). GO and KEGG analyses showed that nitrate reductase activity, nitrate assimilation, and N metabolism were significantly enriched in both the roots and shoots. Transcription factor (TF) and protein kinase analysis showed that genes such as MYB-related (38/38 genes) may function in a tissue-specific manner to respond to LN stress. Moreover, 20 out of 107 N signaling homologous genes were differentially expressed in wheat. A total of 47 transcriptome datasets were used for weighted gene co-expression network analysis (17,840 genes), and five TFs were identified as the potential hub regulatory genes involved in the response to LN stress in wheat. Our findings provide insight into the functional mechanisms in response to LN stress and five candidate regulatory genes in wheat. These results will provide a basis for further research on promoting NUE in wheat.

Screening key sorghum germplasms for low-nitrogen tolerance at the seedling stage and identifying from the carbon and nitrogen metabolism.

Sorghum (Sorghum bicolor L.) can withstand drought and heat stress and efficiently utilize water and nutrients. However, the underlying mechanism of its tolerance to low-nitrogen (N) stress remains poorly understood. This study assessed low-N tolerance in 100 sorghum-inbred lines and identified those with exceptional resilience. Principal component analysis, Pearson's correlation, and Y value analysis were used to examine various seedling growth metrics, including plant and root dimensions, biomass, chlorophyll content, root N content, shoot N content, and root/shoot ratio. The genotypes were categorized into four distinct groups based on their respective Y values, revealing a spectrum from highly tolerant to sensitive. Low-N-tolerant sorghum lines maintained higher photosynthetic rates and exhibited increased enzymatic activities linked to carbon and N metabolism in the leaves and roots. Furthermore, low-N-tolerant genotypes had higher levels of key amino acids, including cystine, glycine, histidine, isoleucine, leucine, phenylalanine, threonine, and tyrosine, indicating a robust internal metabolic response to N deficiency. This study provides a comprehensive and reliable approach for the evaluation of sorghum tolerance to low-N environments, sheds light on its morphological and physiological adaptations, and provides valuable insights for future breeding programs and agricultural practices.

Genome-Wide Identification, Characterization, and Expression Analysis of the Amino Acid Permease Gene Family in Soybean

Amino acid permeases (AAPs) play important roles in transporting amino acids in plant species, leading to increased low-nitrogen tolerance, grain yield, or protein content. However, very few AAPs have been characterized in soybean (Glycine max). In this study, we scanned the soybean reference genome and identified a total of 36 AAP genes (named GmAAP). The GmAAPs were phylogenetically divided into three evolutionary clades, with the genes in the same clades sharing similar gene structures and domain organization. We also showed that seventeen GmAAP genes on ten chromosomes were in collinearity, likely due to whole-genome duplication. Further analysis revealed a variety of cis-acting regulatory elements (such as hormone response elements (ABRE, ERE, GARE, P-box, and TGA-element), stress response elements (LTR, MBS, MYB-related components, TC-rich repeats, TCA-element, and WUN-motif), the tissue expression element (GCN4-motif), and the circadian regulatory element (circadian) present in the 2 kb region of the GmAAP promoter region, demonstrating functional diversity and expression specificity. RNA-Seq data and quantitative real-time PCR identified five GmAAPs showing differential expression under nitrogen limitation, including GmAAP3, GmAAP5, and GmAAP8 showing downregulation while GmAAP14, GmAAP29 showed upregulation, suggesting their involvement in low-nitrogen stress response. These results provide comprehensive information on soybean AAP genes in nitrogen stress, and provide putative candidates with possible roles in enhancing amino acid delivery to seeds for yield improvement.

Nickel Ions Enhanced the Adaptability of Tomato Seedling Roots to Low-Nitrogen Stress by Improving Their Antioxidant Capacity

To elucidate the physiological mechanisms underlying the impact of exogenous nickel ions (Ni2+) on the adaptability of tomato (Solanum lycopersicum L.) seedling roots to low-nitrogen levels, the cultivar ‘Micro Tom’ was selected as the experimental material and cultivated hydroponically in the cultivation room of the Fujian Agriculture and Forestry University. Two distinct nitrogen concentrations (7.66 and 0.383 mmol·L−1) and two different levels of Ni2+ (0 and 0.1 mg·L−1 of NiSO4·6H2O) were employed as treatments. On the 9th day of cultivation, we measured the root biomass, the concentrations of antioxidant compounds, and the activities of antioxidant enzymes in the tomato seedlings. The study showed that when the nitrogen levels were low, the growth and development of the tomato seedling roots were hindered. This led to a significant increase in the levels of hydrogen peroxide (H2O2), superoxide anion (O2−), and malondialdehyde (MDA), indicating oxidative damage to the roots. Conversely, treatment with Ni2+ induced a notable increase in the activity of antioxidant enzymes in the seedlings and augmented the accumulation of nonenzymatic antioxidants, such as ascorbic acid (ASA) and reduced glutathione (GSH), thereby enhancing the operational efficiency of the ascorbate–glutathione cycle (ASA–GSH). Consequently, this led to substantial reductions in the H2O2 and MDA levels, ultimately mitigating the oxidative damage inflicted on the tomato seedling roots subjected to low-nitrogen stress. In conclusion, exogenous Ni2+ can reduce the peroxidative damage of tomato seedlings by promoting antioxidase activity in tomato seedlings under low-nitrogen stress, improve the tolerance of tomato seedlings to low-nitrogen stress, and maintain the normal growth and development of tomato seedlings.

Brassinosteroids alleviates wheat floret degeneration under low nitrogen stress by promoting the distribution of sucrose from stems to spikes

The trade-off between yield and environmental effects caused by nitrogen fertilizer application is an important issue in wheat production. A reduction in fertile florets is one of the main reasons for decreased yield under low nitrogen application rate. Brassinosteroids (BRs) have been found to play a role in nitrogen-induced rice spikelet degeneration. However, whether BRs play a role in wheat floret development and the mechanisms involved are not clear. Therefore, different nitrogen gradient experiment and exogenous spraying experiment were conducted to investigate the role and mechanism of BRs in wheat floret development under low nitrogen stress. The results showed that as the nitrogen application decreased, the endogenous BRs content of spikes decreased, photosynthesis weakened, and total carbon, soluble sugar and starch of spikes decreased, leading to a reduction in the number of fertile florets. Under low nitrogen stress, exogenous spraying of 24-epibrassinolide promoted photosynthesis, stimulated stem fructan hydrolysis and the utilization and storage of sucrose in spikes, directing more carbohydrates to the spikes and increasing the number of fertile florets. In conclusion, BRs mediate the effects of nitrogen fertilizer on wheat floret development, and under low nitrogen stress, foliar spraying of 24-epibrassinolide promotes the flow of carbohydrates from the stem to the spikes, alleviating wheat floret degeneration.

Physiological and Transcriptomic Analysis Provides Insights into Low Nitrogen Stress in Foxtail Millet (Setaria italica L.).

Foxtail millet (Setaria italica (L.) P. Beauv) is an important food and forage crop that is well adapted to nutrient-poor soils. However, our understanding of how different LN-tolerant foxtail millet varieties adapt to long-term low nitrogen (LN) stress at the physiological and molecular levels remains limited. In this study, two foxtail millet varieties with contrasting LN tolerance properties were investigated through analyses of physiological parameters and transcriptomics. The physiological results indicate that JG20 (high tolerance to LN) exhibited superior biomass accumulation both in its shoots and roots, and higher nitrogen content, soluble sugar concentration, soluble protein concentration, zeatin concentration in shoot, and lower soluble sugar and soluble protein concentration in its roots compared to JG22 (sensitive to LN) under LN, this indicated that the LN-tolerant foxtail millet variety can allocate more functional substance to its shoots to sustain aboveground growth and maintain high root activity by utilizing low soluble sugar and protein under LN conditions. In the transcriptomics analysis, JG20 exhibited a greater number of differentially expressed genes (DEGs) compared to JG22 in both its shoots and roots in response to LN stress. These LN-responsive genes were enriched in glycolysis metabolism, photosynthesis, hormone metabolism, and nitrogen metabolism. Furthermore, in the shoots, the glutamine synthetase gene SiGS5, chlorophyll apoprotein of photosystem II gene SiPsbQ, ATP synthase subunit gene Sib, zeatin synthesis genes SiAHP1, and aldose 1-epimerase gene SiAEP, and, in the roots, the high-affinity nitrate transporter genes SiNRT2.3, SiNRT2.4, glutamate synthase gene SiGOGAT2, fructose-bisphosphate aldolase gene SiFBA5, were important genes involved in the LN tolerance of the foxtail millet variety. Hence, our study implies that the identified genes and metabolic pathways contribute valuable insights into the mechanisms underlying LN tolerance in foxtail millet.

Overexpression of Cytosolic Glyceraldehyde-3-Phosphate Dehydrogenase 1 Gene Improves Nitrogen Absorption and Utilization in Potato

Nitrogen is one of the most important elements for improving potato yield. However, excessive application of nitrogenous fertilizer not only produces river and other environmental pollution but also increases agricultural production costs. In recent years, to explain the molecular mechanisms of nitrogen metabolites, some vital genes involved have been reported; however, only limited success has been achieved in potato. Here, we report that the expression of cytosolic glyceraldehyde-3-phosphate dehydrogenase 1 (StGAPC1) is increased under low-nitrogen stress. StGAPC1-overexpressing potato seedlings had more biomass and a significant increase in total nitrogen content and root nitrate influx rate compared to the wild type. The overexpression of StGAPC1 also increased the expression of nitrate transporters and increased ROS system activity to reduce hydrogen peroxide content under low-nitrogen stress. Our results provide a foundation for further research on StGAPC1 function in nitrogen absorption and utilization mechanisms in potato.

A comparative analysis of root traits in diverse wheat seedlings screened under optimum and low nitrogen stress in hydroponics

A comparative analysis of root traits in diverse wheat seedlings screened under optimum and low nitrogen stress in hydroponics

Different genotypes of tartary buckwheat can regulate the transformation of nitrogen through the secretion of organic acids under low nitrogen stress

Roots respond to nitrogen deficiency by altering their physiological and metabolic responses. Soil nitrogen invertase activity and the diversity of nitrifying microorganisms (AOA and AOB) significantly affect the nitrogen cycle in soil. As an important part of root exudates, organic acids can reflect the dynamic changes of plant roots in soil. However, it is not clear how the barren-tolerant genotype plant varieties show their stress resistance through organic acids regulating nitrogen transformation under low nitrogen environment. Here, we selected two Tartary buckwheat varieties: HeiFeng1 (HF, low nitrogen sensitive cultivar) and DiQing (DQ, low nitrogen tolerance cultivar) for a two-year experiment. The urea addition levels of 0, 80, and 160 mg kg−1 soil were respectively set as nitrogen sources, and ozone sterilization experiments were designed to completely remove microbial effects to distinguish the contribution of plant roots and microorganisms to nitrogen invertase and organic acids (400 ml 1 mol L−1 ozone water daily, 0 nitrogen fertilizer, O3). The results showed that malic acid and tartaric acid were the main organic acids secreted by tartary buckwheat. The total amount and richness of organic acids in Diqing were higher than those in Heifeng, and organic acids directly affected the diversity of ammonia-oxidizing microorganisms (AOA and AOB), changed the soil environment by affecting soil water content, and indirectly affected the activity of nitrogen invertase. At the same time, it was found that the increase of nitrogen application rate would increase the proportion of key bacteria such as Nirtrosomonas and Nitrosospira in AOB of tartary buckwheat soil. Finally, Diqing tartary buckwheat was proved to be more suitable for planting in the nitrogen-deficient Loess Plateau, but attention should be paid to the negative effects of AOB and organic acids.

Mechanisms governing the impact of nitrogen stress on the formation of secondary metabolites in Artemisia argyi leaves

Nitrogen is a key factor in various physiological and metabolic processes in plants. Providing an adequate supply of nitrogen is essential for improving the total yield and quality of the medicinal plant Artemisia argyi (A. argyi), but the underlying mechanisms of how this nutrient alters the crop remains unclear. In this study, we conducted a series of pot experiments to investigate the agronomic traits and active components in the leaves of A. argyi plants under low and high nitrogen stress. Additionally, we used transcriptome analysis and RT-qPCR to explore the molecular pathways associated with nitrogen stress. Our results demonstrate a dramatic increase in the accumulation of phenolic acids and flavonoids in the low nitrogen (LN) stress group compared to the control (CK), with increases of 40.00% and 79.49%, respectively. Interestingly, plants in the high nitrogen (HN) stress group exhibited enhanced plant growth with larger leaves, thicker stems, and a 3% increase in volatile oil content compared to the CK. Moreover, A. argyi in the HN group displayed a 66% increase in volatile oil concentration compared to the LN group. Our combined transcriptome and q-PCR results indicate that LN stress promotes the expression of genes involved in flavonoid synthesis, while HN stress promotes the expression of genes related to terpene skeleton production and photosynthesis. Taken together, these findings suggest that different gene expression levels under LN and HN stress contribute to the photosynthesis capacity and the accumulation of active ingredients in A. argyi leaves. Our results elucidate the physiological and molecular mechanisms of nitrogen stress on A. argyi secondary metabolites and guide fertilization strategies for plant cultivation.

Ethylene Regulates Combined Drought and Low Nitrogen Tolerance in Wheat: Proteomic Analysis

Wheat is a staple crop in China’s arid and semi-arid regions. Drought and low nitrogen (LN) are two major constraints to wheat growth and production. However, the molecular mechanism underlying wheat response to both drought and LN stress remains unknown. Accordingly, we conducted a proteomic study on the roots of two wheat varieties, Chang6878 (drought tolerant) and Shi4185 (drought sensitive) and compared the differences between drought and combined drought and LN stress treatments. In total, 5143 proteins were identified, of which 163 differentially abundant proteins (DAPs) were uniquely upregulated under drought and LN stress in Chang6878. Enrichment analysis showed that DAPs were mainly involved in mitogen-activated protein kinase signaling, phenylpropanoid biosynthesis, glutathione metabolism, ethylene biosynthesis, ethylene signal transduction, and oxidation–reduction reactions. These DAPs were verified via parallel reaction monitoring and quantitative real-time polymerase chain reaction. Chang6878 was treated with the ethylene synthesis precursor 1-aminocyclopropanecarboxylic acid, and its resistance to drought and LN stress improved. After treatment with the ethylene synthesis inhibitor silver nitrate and ethylene signal transduction inhibitor 1-methylcyclopropene, drought and LN stress resistance reduced. These results provide novel insights into the tolerance mechanisms of Chang6878 to drought and LN stress by altering ethylene synthesis and signal transduction. This study provides a reference for breeding drought- and low-nitrogen-tolerant wheat germplasm resources and a theoretical basis for maintaining food security in arid, barren areas.

Conditioning to Low-Nitrogen Regimes Results in Rice Plants Better Adapted to Low-Nitrogen Stress Through DNA Methylation, Metabolic, and Morpho-physiological Modifications

Conditioning to Low-Nitrogen Regimes Results in Rice Plants Better Adapted to Low-Nitrogen Stress Through DNA Methylation, Metabolic, and Morpho-physiological Modifications

Overexpression of PsAMT1.2 in poplar enhances nitrogen utilization and resistance to drought stress.

Ammonium is an important form of inorganic nitrogen, which is essential for plant growth and development, and the uptake of ammonium is mediated by different members of ammonium transporters (AMTs). It is reported that PsAMT1.2 is specially expressed in the root of poplar, and the overexpression of PsAMT1.2 could improve plant growth and the salt tolerance of poplar. However, the role of AMTs in plant drought and low nitrogen (LN) resistance remains unclear. To understand the role of PsAMT1.2 in drought and LN tolerance, the response of PsAMT1.2-overexpression poplar to polyethylene glycol (PEG)-simulated drought stress (5% PEG) under LN (0.001mM NH4NO3) and moderate nitrogen (0.5mM NH4NO3) conditions was investigated. The PsAMT1.2-overexpression poplar showed better growth with increased stem increment, net photosynthetic rate, chlorophyll content, root length, root area, average root diameter and root volume under drought and/or LN stress compared with the wild type (WT). Meanwhile, the content of malondialdehyde significantly decreased, and the activities of superoxide dismutase and catalase significantly increased in the roots and leaves of PsAMT1.2-overexpression poplar compared with WT. The content of NH4+ and NO2- in the roots and leaves of PsAMT1.2-overexpression poplar was increased, and nitrogen metabolism-related genes, such as GS1.3, GS2, Fd-GOGAT and NADH-GOGAT, were significantly upregulated in the roots and/or leaves of PsAMT1.2-overexpression poplar compared with WT under drought and LN stress. The result of this study would be helpful for understanding the function of PsAMT1.2 in plant drought and LN tolerance and also provides a new insight into improving the drought and LN tolerance of Populus at the molecular level.