2-oxoglutarate Aminotransferase Research Articles

Post-anthesis dry matter and nitrogen accumulation, partitioning, and translocation in maize under different nitrate-ammonium ratios in Northwestern China.

An appropriate supply of ammonium (NH4+) in addition to nitrate (NO3-) can greatly improve plant growth and promote maize productivity. However, knowledge gaps exist regarding the mechanisms by which different nitrogen (N) fertilizer sources affect the enzymatic activity of nitrogen metabolism and non-structural carbohydrates during the post-anthesis period. A field experiment across 3-year was carried out to explore the effects of four nitrateammonium ratio (NO3-/NH4+ = 1:0 (N1), 1:1 (N2), 1:3 (N3), and 3:1 (N4)) on postanthesis dry matter (DM) and N accumulation, partitioning, transportation, and grain yield in maize. NO3-/NH4+ ratio with 3:1 improved the enzymatic activity of N metabolism and non-structural carbohydrate accumulation, which strongly promoted the transfer of DM and N in vegetative organs to reproductive organs and improved the pre-anthesis DM and nitrogen translocation efficiency. The enzymatic activities of nitrate reductase, nitrite reductase, glutamine synthetase, glutamine oxoglutarate aminotransferase, and non-structural carbohydrate accumulation under N4 treatment were increased by 9.30%-32.82%, 13.19%-37.94%, 4.11%-16.00%, 11.19%-30.82%, and 14.89%-31.71% compared with the other treatments. Mixed NO3--N and NH4+-N increased the total DM accumulation at the anthesis and maturity stages, simultaneously decreasing the DM partitioning of stem, increasing total DM, DM translocation efficiency (DMtE), and contribution of pre-anthesis assimilates to the grain (CAPG) in 2015 and 2017, promoting the transfer of DM from stem to grain. Furthermore, the grain yield increased by 3.31%-9.94% (2015), 68.6%-26.30% (2016), and 8.292%-36.08% (2017) under the N4 treatment compared to the N1, N2, and N3 treatments. The study showed that a NO3-/NH4+ ratio of 3:1 is recommended for high-yield and sustainable maize management strategies in Northwestern China.

Application of multi-omic features clustering and pathway enrichment to clarify the impact of vitamin B2 supplementation on broiler caeca microbiome.

The results of omic methodologies are often reported as separate datasets. In this study we applied for the first time multi-omic features clustering and pathway enrichment to clarify the biological impact of vitamin B2 supplementation on broiler caeca microbiome. The caeca contents of broilers fed +50 and +100 mg/kg vitamin B2 were analyzed by shotgun metagenomic and metabolomic. Latent variables extracted from NMR spectra, as well as taxonomic and functional features profiled from metagenomes, were integrated to characterize the effect of vitamin B2 in modulating caeca microbiome. A pathway-based network was obtained by mapping the observed input genes and compounds, highlighting connected strands of metabolic ways through pathway-enrichment analysis. At day 14, the taxonomic, functional and metabolomic features in the caeca of tested broilers showed some degree of separation between control and treated groups, becoming fully clear at 28 days and persisting up to 42 days. In the caeca of birds belonging to the control group Alistipes spp. was the signature species, while the signature species in the caeca of broilers fed +50 and +100 mg/kg vitamin B2 were Bacteroides fragilis and Lactobacillus crispatus, Lactobacillus reuteri, Ruminococcus torques, Subdoligranum spp., respectively. The pathway enrichment analysis highlighted that the specific biochemical pathways enhanced by the supplementations of vitamin B2 were N-Formyl-L-aspartate amidohydrolase, producing Aspartate and Formate; L-Alanine:2-oxoglutarate amino transferase, supporting the conversion of L-Alanine and 2-Oxoglutarate in Pyruvate and L-Glutamate; 1D-myo-inositol 1/4 phosphate phosphohydrolase, converting Inositol 1/4-phosphate and water in myo-Inositol and Orthophosphate. The results of this study demonstrated that the caeca of birds fed +50 and + 100 mg/kg were those characterized by taxonomic groups more beneficial to the host and with a higher concentration of myo-inositol, formic acid, amino acids and pyruvate involved in glycolysis and amino acid biosynthesis. In this study we demonstrated how to perform multi-omic features integration to describe the biochemical mechanisms enhanced by the supplementation of different concentrations of vitamin B2 in the poultry diet. The relationship between vitamin B2 supplementation and myo-inositol production was highlighted in our study for the first time.

The rice NUCLEAR FACTOR-YA5 and MICRORNA169a module promotes nitrogen utilization during nitrogen deficiency.

Nitrogen (N) is essential for plant growth and development. Therefore, understanding its utilization is essential for improving crop productivity. However, much remains to be learned about plant N sensing and signaling. Here, rice (Oryza sativa) NUCLEAR FACTOR-YA5 (OsNF-YA5) expression was tightly regulated by N status and induced under N-deficient conditions. Overexpression (OE) of OsNF-YA5 in rice resulted in increased chlorophyll levels and delayed senescence compared to control plants under normal N conditions. Agronomic traits were significantly improved in OE plants and impaired in knockout mutants under N-deficient conditions. Using a dexamethasone-inducible system, we identified the putative targets of OsNF-YA5 that include amino acid, nitrate/peptide transporters, and NITRATE TRANSPORTER 1.1A (OsNRT1.1A), which functions as a key transporter in rice. OsNF-YA5 directly enhanced OsNRT1.1A expression and N uptake rate under N-deficient conditions. Besides, overexpression of OsNF-YA5 also enhanced the expression of GLUTAMINE SYNTHETASE 1/2 (GS1/2) and GLUTAMINE OXOGLUTARATE AMINOTRANSFERASE 1/2 (GOGAT1/2), increasing free amino acid contents under N-deficient conditions. Osa-miR169a expression showed an opposite pattern with OsNF-YA5 depending on N status. Further analysis revealed that osa-miR169a negatively regulates OsNF-YA5 expression and N utilization, demonstrating that an OsNF-YA5/osa-miR169a module tightly regulates rice N utilization for adaptation to N status.

Effects of Biological Nitrogen Metabolism on Glufosinate-Susceptible and -Resistant Goosegrass (Eleusine indica L.).

Glufosinate is a broad-spectrum herbicide used to control most weeds in agriculture worldwide. Goosegrass (Eleusine indica L.) is one of the top ten malignant weeds across the world, showing high tolerance to glufosinate via different mechanisms that are not yet fully understood. This study revealed that nitrogen metabolism could be a target-resistant site, providing clues to finally clarify the mechanism of glufosinate resistance in resistant goosegrass populations. Compared to susceptible goosegrass (NX), the resistant goosegrass (AUS and CS) regarding the stress of glufosinate showed stronger resistance with lower ammonia contents, higher target enzyme GS (glutamine synthetase) activity, and lower GOGAT (glutamine 2-oxoglutarate aminotransferase) activity. The GDH (glutamate dehydrogenase) activity of another pathway increased, but its gene expression was downregulated in resistant goosegrass (AUS). Analyzing the transcriptome and proteome data of goosegrass under glufosinate stress at 36 h showed that the KEGG pathway of the nitrogen metabolism was enriched in glufosinate-susceptible goosegrass (NX), but not in glufosinate-resistant goosegrass (CS and AUS). Several putative target genes involved in glufosinate stress countermeasures were identified. This study provides specific insights into the nitrogen metabolism of resistant goosegrass, and gives a basis for future functional verification of glufosinate-tolerance genes in plants.

Chitosan nanoparticles upregulate C and N metabolism in soybean plants grown under elevated levels of atmospheric carbon dioxide

Despite the wide utilization of chitosan nanoparticles (CSNPs) as a promising approach for sustainable agriculture, their efficiency under elevated CO2 (eCO2), has not been evaluated. The interactive effects of CSNPs and eCO2 were evaluated on the growth and C and N metabolism of soybean plants. Plants were treated with CSNPs and grown under ambient CO2 (410 ppm, aCO2) or eCO2 (645 ppm). Regardless of CO2 level, CSNPs improved the net photosynthetic rate. CSNPs aggravated the effect of eCO2 treatment on the levels of non-structural carbohydrates (i.e., glucose, fructose, sucrose, and starch), especially in shoots, which was inconsistence with the upregulation of carbohydrates metabolizing enzymes. Being the most pivotal energetic and signaling organic compounds in higher plants, the synergistic action of CSNPs and eCO2 on the accumulation of soluble sugars upregulated the N metabolism as indicated by induced activities of nitrate reductase, arginase, glutamate dehydrogenase, glutamine synthetase, and glutamine oxoglutarate aminotransferase which was manifested finally as increased shoot and root total nitrogen content as well as proline and aspartate in roots. At the hormonal level, the coexistence of eCO2 with CSNPs further supports their positive impact on the contents of IAA and, to a lesser extent, GAs. The present data prove that the biofertilization capacity of CSNPs is even more potent under futuristic eCO2 levels and could even further improve the growth and resilience of plants.

Improving acetoin production through construction of a genome-scale metabolic model

Acetoin was widely used in food, medicine, and other industries, because of its unique fragrance. Bacillus amyloliquefaciens was recognized as a safe strain and a promising acetoin producer in fermentation. However, due to the complexity of its metabolic network, it had not been fully utilized. Therefore, a genome-scale metabolic network model (iJYQ746) of B. amyloliquefaciens was constructed in this study, containing 746 genes, 1736 reactions, and 1611 metabolites. The results showed that Mg2+, Mn2+, and Fe2+ have inhibitory effects on acetoin. When the stirring speed was 400 rpm, the maximum titer was 49.8 g L−1. Minimization of metabolic adjustments (MOMA) was used to identify potential metabolic modification targets 2-oxoglutarate aminotransferase (serC, EC 2.6.1.52) and glucose-6-phosphate isomerase (pgi, EC 5.3.1.9). These targets could effectively accumulate acetoin by increasing pyruvate content, and the acetoin synthesis rate was increased by 610% and 10%, respectively. This provides a theoretical basis for metabolic engineering to reasonably transform B. amyloliquefaciens and produce acetoin.

Coregulation of glutamine synthetase1;2 (GLN1;2) and NADH-dependent glutamate synthase (GLT1) gene expression in Arabidopsis roots in response to ammonium supply.

Ammonium absorbed by roots is assimilated into amino acids. The glutamine synthetase/glutamate synthase (glutamine 2-oxoglutarate aminotransferase) (GS/GOGAT) cycle is essential to this biological process. In Arabidopsis thaliana, GLN1;2 and GLT1 are the GS and GOGAT isoenzymes induced in response to ammonium supply and playing key roles in ammonium utilization. Although recent studies suggest gene regulatory networks involved in transcriptional regulation of ammonium-responsive genes, direct regulatory mechanisms for ammonium-induced expression of GS/GOGAT remain unclear. In this study, we revealed that the expression of GLN1;2 and GLT1 in Arabidopsis is not directly induced by ammonium but is regulated by glutamine or post-glutamine metabolites produced by ammonium assimilation. Previously, we identified a promoter region required for ammonium-responsive expression of GLN1;2. In this study, we further dissected the ammonium-responsive region of the GLN1;2 promoter and also performed a deletion analysis of the GLT1 promoter, which led to the identification of a conserved ammonium-responsive region. Yeast one-hybrid screening using the ammonium-responsive region of the GLN1;2 promoter as a decoy sequence revealed a trihelix family transcription factor DF1 that binds to this region. A putative DF1 binding site was also found in the ammonium-responsive region of the GLT1 promoter.

Comparative Transcriptomic Analysis Reveals the Negative Response Mechanism of Peanut Root Morphology and Nitrate Assimilation to Nitrogen Deficiency

Root architecture plays a fundamental role in crop yield, which is sensitive to nitrogen fertilizer. Although it is well studied that nitrogen fertilizer significantly promotes peanut (Arachis hypogaea L.) growth and yield, less information was available on how its root development responds to nitrogen deficiency. In this study, the growth and development of roots were inhibited, as indicated by the significantly decreased root dry weight and length and the lateral root number, especially under 10 days of nitrogen deficiency treatment. The activities and the expression of the genes related to nitrogen assimilation enzymes including nitrate reductase, glutamine synthetase, glutamate dehydrogenase, and glutamine oxoglutarate aminotransferase and the genes encoding the nitrate transporters were significantly decreased under 10 days of nitrogen deficiency treatment, which may lead to a decrease in nitrate content, as indicated by the significantly decreased nitrogen balance index. Transcriptome sequencing revealed a total of 293 (119 up- and 174 downregulated) and 2271 (1165 up- and 1106 downregulated) differentially expressed genes (DEGs) identified after five and ten days of nitrogen deficiency treatments, respectively. Bioinformatic analysis showed that these DEGs were mainly involved in nitrate transportation and assimilation, phytohormone signal transduction, and the lignin biosynthesis pathway. Furthermore, a putative schematic diagram of nitrogen deficiency inhibiting root growth was established, which gives us a better understanding of nitrogen metabolism in peanut roots and a theoretical basis for improving nitrogen use efficiency.

Effect of herbicide stress on synchronization of carbon and nitrogen metabolism in lentil (Lens culinaris Medik.)

Weed invasion causes significant yield losses in lentil. Imazethapyr (IM), a broad-spectrum herbicide inhibits the biosynthesis of branched chain amino acids necessary for plant growth. Plant growth depends upon translocation of photo-assimilates and their partitioning regulated by carbon and nitrogen metabolism. This study aimed to investigate the impact of imazethapyr spray on carbon and nitrogen metabolism in tolerant (LL1397 and LL1612) and susceptible (FLIP2004-7L and PL07) lentil genotypes during vegetative and reproductive development. Significantly higher activities of invertases and sucrose synthase (cleavage) in leaves and in podwall and seeds during early phase of development in tolerant genotypes were observed as compared to susceptible genotypes under herbicide stress that might be responsible for providing hexoses required for their growth. Activities of sucrose synthesizing enzymes, sucrose phosphate synthase and sucrose synthase (synthesis) increased significantly in podwalls and seeds of LL1397 and LL1612 genotypes during later phase of development towards maturity while the activities decreased in FLIP2004-7L and PL07 genotypes under herbicide stress. Activities of nitrate and nitrite reductase, glutamine 2-oxoglutarate aminotransferase, glutamine synthetase and glutamate dehydrogenase were increased in leaves, podwalls and seeds of LL1397 and LL1612 under herbicide stress. A proper synchronization of carbon and nitrogen metabolism in tolerant lentil genotypes during vegetative and reproductive phase might be one of the mechanisms for their recovery from herbicide stress. This first ever comprehensive information will provide a basis for future studies on the molecular mechanism of source sink relationship in lentil under herbicide stress and will be utilized in breeding programmes.

Photoautotrophic Growth Rate Enhancement of Synechocystis sp. PCC6803 by Heterologous Production of 2-Oxoglutarate:Ferredoxin Oxidoreductase from Chlorobaculum tepidum.

2-Oxoglutarate:ferredoxin oxidoreductase from Chlorobaculum tepidum (CtOGOR) is a carbon-fixing enzyme in the reductive TCA cycle that reversibly carboxylates succinyl-CoA to yield 2-oxoglutarate. CtOGOR is a heterotetramer of two large (α = 68 kDa) and two small (β = 38 kDa) subunits. The αβ protomer harbors one thiamine pyrophosphate and two 4Fe-4S clusters. Nonetheless, the enzyme has a considerable oxygen tolerance with a half-life of 143 min at 215 μM dissolved oxygen. Kinetic analyses of the purified recombinant CtOGOR revealed a lower Km for succinyl-CoA than for 2-oxoglutarate. Cellular levels of 2-oxoglutarate and glutamate-a product of glutamine oxoglutarate aminotransferase and glutamate dehydrogenase-increased more than twofold in the exponential phase compared with the control strain, leading to an approximately >30% increase in the photoautotrophic growth rate. Thus, CtOGOR was successfully produced in Synechocystis, thereby boosting carboxylation, resulting in enhanced photoautotrophic growth.

Effects of bisphenol A on antioxidation and nitrogen assimilation of maize seedlings roots

Bisphenol A (BPA) is becoming a potential environmental toxicity factor. However, BPA's effect and function mechanism on maize roots remain unknown. Here, we investigated characters of root growth of maize seedlings exposed to BPA for 8 d and without BPA for 3 d, and a series of indicators on reactive oxygen homeostasis and nitrogen assimilation were measured. High-dose BPA(15 and 50 mg·L−1) suppressed the root growth and caused increased contents of O2ˑ-, H2O2 and MDA in maize seedling roots. The disturbed ROS homeostasis resulted from the change of antioxidant enzymes, including the increase of APX, GPX, and CAT, and decrease of SOD and POD, and a decrease of antioxidant substance GSH. Meanwhile, High-dose BPA caused a decrease in the soluble protein content, nitrate reductase (NR), glutamate dehydrogenase (GDH), and glutamine oxoglutarate aminotransferase (GOGAT) under the BPA processing phase and recovery period. The low-dose BPA（1.5 and 5 mg·L−1）significantly promoted root growth of maize seedlings and maintained the ROS homeostasis through antioxidant enzyme APX and GPX eliminating redundant ROS. Our results showed that BPA could cause a dual effect on the root growth of maize seedlings, that is, promotion of low-dose and inhibition of high-dose, through ROS homeostasis and nitrogen assimilation in Zea mays.

MeNPF4.5 Improves Cassava Nitrogen Use Efficiency and Yield by Regulating Nitrogen Uptake and Allocation.

Improving nitrogen use efficiency (NUE) is a very important goal of crop breeding throughout the world. Cassava is an important food and energy crop in tropical and subtropical regions, and it mainly use nitrate as an N source. To evaluate the effect of the nitrate transporter gene MeNPF4.5 on the uptake and utilization of N in cassava, two MeNPF4.5 overexpression lines (MeNPF4.5 OE-22 and MeNPF4.5 OE-34) and one MeNPF4.5 RNA interference (RNAi) line (MeNPF4.5 Ri-1) were used for a tissue culture experiment, combining with a field trial. The results indicated that MeNPF4.5 is a plasma membrane transporter mainly expressed in roots. The gene is induced by NO3–. Compared with the wild type, MeNPF4.5 OE-22 exhibited improved growth, yield, and NUE under both low N and normal N levels, especially in the normal N treatment. However, the growth and N uptake of RNAi plants were significantly reduced, indicating poor N uptake and utilization capacity. In addition, photosynthesis and the activities of N metabolism-related enzymes (glutamine synthetase, glutamine oxoglutarate aminotransferase, and glutamate dehydrogenase) of leaves in overexpression lines were significantly higher than those in wild type. Interestingly, the RNAi line increased enzymatic activity but decreased photosynthesis. IAA content of roots in overexpressed lines were lower than that in wild type under low N level, but higher than that of wild type under normal N level. The RNAi line increased IAA content of roots under both N levels. The IAA content of leaves in the overexpression lines was significantly higher than that of the wild type, but showed negative effects on that of the RNAi lines. Thus, our results demonstrated that the MeNPF4.5 nitrate transporter is involved in regulating the uptake and utilization of N in cassava, which leads to the increase of N metabolizing enzyme activity and photosynthesis, along with the change of endogenous hormones, thereby improving the NUE and yield of cassava. These findings shed light that MeNPF4.5 is involved in N use efficiency use in cassava.

Rhizobium Symbiosis Leads to Increased Drought Tolerance in Chinese Milk Vetch (Astragalus sinicus L.)

Chinese milk vetch (Astragalus sinicus L.) is an important leguminous green manure that frequently suffers from seasonal drought. To improve its drought tolerance, the effects of rhizobia inoculation on drought tolerance and the underlying physiological mechanism were investigated. Drought tolerance in combination with nitrogen assimilation, free amino acids, and polyamines was investigated in milk vetch with active nodules (AN), with inactive nodules (LN), or without nodules (NN). AN plants had increased drought tolerance compared to LN and NN plants. Glutamine synthetase (GS), glutamine 2-oxoglutarate amino transferase (GOGAT), and glutamate dehydrogenase (GDH) activities were decreased after drought, with higher levels in AN plants than in LN and NN plants under both control and drought stress conditions. Higher levels of proline (Pro), arginine (Ala), alanine (Ala), and glutamate (Glu) were observed in AN plants compared with LN and NN plants. Putrescine (Put), spermidine (Spd), and spermine (Spm) levels were increased in response to drought, and higher levels of Put and Spd were maintained in AN plants. It is suggested that active nodulation leads to increased drought tolerance in milk vetch, which is associated with improved nitrogen fixation and ammonium assimilation, which in turn lead to the promotion of free amino-acid and polyamine synthesis.

Regulation of nitrogen starvation responses by the alarmone (p)ppGpp in rice

Nitrogen is an essential macronutrient for all living organisms and is critical for crop productivity and quality. In higher plants, inorganic nitrogen is absorbed through roots and then assimilated into amino acids by the highly conserved glutamine synthetase/glutamine:2-oxoglutarate aminotransferase (GS/GOGAT) cycle. How nitrogen metabolism and nitrogen starvation responses of plants are regulated remains largely unknown. Previous studies revealed that mutations in the rice ABNORMAL CYTOKININ RESPONSE1 (ABC1) gene encoding Fd-GOGAT cause a typical nitrogen deficiency syndrome. Here, we show that ARE2 (for ABC1 REPRESSOR2) is a key regulator of nitrogen starvation responses in rice. The are2 mutations partially rescue the nitrogen-deficient phenotype of abc1 and the are2 mutants show enhanced tolerance to nitrogen deficiency, suggesting that ARE2 genetically interacts with ABC1/Fd-GOGAT. ARE2 encodes a chloroplast-localized RelA/SpoT homolog protein that catalyzes the hydrolysis of guanosine pentaphosphate or tetraphosphate (p)ppGpp, an alarmone regulating the stringent response in bacteria under nutritional stress conditions. The are2 mutants accumulate excessive amounts of (p)ppGpp, which correlate with lower levels of photosynthetic proteins and higher amino acid levels. Collectively, these observations suggest that the alarmone (p)ppGpp mediates nitrogen stress responses and may constitute a highly conserved mechanism from bacteria to plants.

Effects of Corn Intercropping with Soybean/Peanut/Millet on the Biomass and Yield of Corn under Fertilizer Reduction

Corn (Zea mays L.) is one of the key grain crops in China. In fields, the two crops of soybean (Glycine max L.) and peanut (Arachis hypogaea L.), which have nitrogen-fixing capacity (NFC), are generally used to intercrop with corn to improve plant physiology and production ability of corn even under fertilizer reduction. To explore a more scientific and reasonable way to plant corn, and simultaneously reduce the use of chemical fertilizers and pesticides, the impacts of corn intercropping with two NFC crops (including soybean and peanut) and the a non-NFC crop (i.e., millet (Setaria italica)) through five planting patterns, including three intercropping patterns (2 corn rows to 2, 3, and 4 NFC-crop rows or 2, 4, and 6 millet rows) and two sole crop patterns of corn and soybean, peanut, or millet under normal (600 kg/ha) and reduced (375 kg/ha) levels of NPK (N:P2O5:K2O = 15:15:15) fertilization levels on the activity of N-metabolism-related enzymes in corn rhizosphere soil and corn leaves, and plant biomass and yield of corn were researched in this study. The results showed that fertilizer reduction significantly decreased the plant biomass and grain yield of the sole crop corn. The intercropping type and planting pattern both had significant effects on the activities of N-metabolism-related enzyme of soil alkaline protease (S-ALPT), and glutamine oxoglutarate aminotransferase (GOGAT), glutamate synthetase (GS), and nitrate reductase (NR) in the leaves of corn plants. The intercropping type of corn with soybean through the planting pattern of 2 corn rows to 4 soybean rows significantly improved the activities of N-metabolism-related enzymes in soil and corn leaves even under the fertilizer reduction. The intercropping pattern of corn-soybean was the most beneficial to increase the total nitrogen content in soil and corn leaves. In addition, the intercropping significantly increased the soil microbial diversity under normal fertilizer. Furthermore, fertilizer reduction significantly increased soil microbial diversity of the corn sole crop. Therefore, it is concluded that for corn in intercropping systems, the best and the worst companion crop were, respectively, soybean and millet.

Nitrogen Assimilation Related Genes in Brassicanapus: Systematic Characterization and Expression Analysis Identified Hub Genes in Multiple Nutrient Stress Responses.

Nitrogen (N) is an essential macronutrient for plants. However, little is known about the molecular regulation of N assimilation in Brassica napus, one of the most important oil crops worldwide. Here, we carried out a comprehensive genome-wide analysis of the N assimilation related genes (NAGs) in B. napus. A total of 67 NAGs were identified encoding major enzymes involved in N assimilation, including asparagine synthetase (AS), glutamate dehydrogenase (GDH), glutamine oxoglutarate aminotransferase (GOGAT), glutamine synthetase (GS), nitrite reductase (NiR), nitrate reductase (NR). The syntenic analysis revealed that segmental duplication and whole-genome duplication were the main expansion pattern during gene evolution. Each NAG family showed different degrees of differentiation in characterization, gene structure, conserved motifs and cis-elements. Furthermore, diverse responses of NAG to multiple nutrient stresses were observed. Among them, more NAGs were regulated by N deficiency and ammonium toxicity than by phosphorus and potassium deprivations. Moreover, 12 hub genes responding to N starvation were identified, which may play vital roles in N utilization. Taken together, our results provide a basis for further functional research of NAGs in rapeseed N assimilation and also put forward new points in their responses to contrasting nutrient stresses.

Genomic identification of nitrogen assimilation-related genes and transcriptional characterization of their responses to nitrogen in allotetraploid rapeseed

Nitrogen (N) is an essential macronutrient to maintain plant growth and development. Plants absorb nitrate-N or ammonium-N in the environment and undergo reduction reactions catalyzed by nitrate reductase (NR), nitrite reductase (NIR), glutamine synthetase (GS), and glutamine oxoglutarate aminotransferase (GOGAT) within plants. A total of 42 N assimilation-related genes (NAG) members were identified in rapeseed. Darwin's evolutionary pressure analysis showed that rapeseed NAGs underwent purification selection. Cis-element analysis revealed differences in the transcriptional regulation of NAGs between Arabidopsis and rapeseed. Expression analyses revealed that NRs were expressed mainly in old leaves, NIRs were expressed mainly in old leaves and lower stem peels, while the expression situation between different subfamilies of GSs and GOGATs was more complicated. Differential expression of NAGs suggested that they might be involved in abiotic stresses. The above results greatly enriched our understanding of NAGs' molecular characteristics and provided central gene resources for NAGs-mediated NUE improvement in rapeseed.

Whole-Genome Duplication Facilitated the Evolution of C4 Photosynthesis in Gynandropsis gynandra.

In higher plants, whole-genome duplication (WGD) is thought to facilitate the evolution of C4 photosynthesis from C3 photosynthesis. To understand this issue, we used new and existing leaf-development transcriptomes to construct two coding sequence databases for C4Gynandropsis gynandra and C3Tarenaya hassleriana, which shared a WGD before their divergence. We compared duplicated genes in the two species and found that the WGD contributed to four aspects of the evolution of C4 photosynthesis in G. gynandra. First, G. gynandra has retained the duplicates of ALAAT (alanine aminotransferase) and GOGAT (glutamine oxoglutarate aminotransferase) for nitrogen recycling to establish a photorespiratory CO2 pump in bundle sheath (BS) cells for increasing photosynthesis efficiency, suggesting that G. gynandra experienced a C3–C4 intermediate stage during the C4 evolution. Second, G. gynandra has retained almost all known vein-development-related paralogous genes derived from the WGD event, likely contributing to the high vein complexity of G. gynandra. Third, the WGD facilitated the evolution of C4 enzyme genes and their recruitment into the C4 pathway. Fourth, several genes encoding photosystem I proteins were derived from the WGD and are upregulated in G. gynandra, likely enabling the NADH dehydrogenase-like complex to produce extra ATPs for the C4 CO2 concentration mechanism. Thus, the WGD apparently played an enabler role in the evolution of C4 photosynthesis in G. gynandra. Importantly, an ALAAT duplicate became highly expressed in BS cells in G. gynandra, facilitating nitrogen recycling and transition to the C4 cycle. This study revealed how WDG may facilitate C4 photosynthesis evolution.

Biochar induced improvement in root system architecture enhances nutrient assimilation by cotton plant seedlings

BackgroundRaising nitrogen use efficiency of crops by improving root system architecture is highly essential not only to reduce costs of agricultural production but also to mitigate climate change. The physiological mechanisms of how biochar affects nitrogen assimilation by crop seedlings have not been well elucidated.ResultsHere, we report changes in root system architecture, activities of the key enzymes involved in nitrogen assimilation, and cytokinin (CTK) at the seedling stage of cotton with reduced urea usage and biochar application at different soil layers (0–10 cm and 10–20 cm). Active root absorption area, fresh weight, and nitrogen agronomic efficiency increased significantly when urea usage was reduced by 25% and biochar was applied in the surface soil layer. Glutamine oxoglutarate amino transferase (GOGAT) activity was closely related to the application depth of urea/biochar, and it increased when urea/biochar was applied in the 0–10 cm layer. Glutamic-pyruvic transaminase activity (GPT) increased significantly as well. Nitrate reductase (NR) activity was stimulated by CTK in the very fine roots but inhibited in the fine roots. In addition, AMT1;1, gdh3, and gdh2 were significantly up-regulated in the very fine roots when urea usage was reduced by 25% and biochar was applied.ConclusionNitrogen assimilation efficiency was significantly affected when urea usage was reduced by 25% and biochar was applied in the surface soil layer at the seedling stage of cotton. The co-expression of gdh3 and gdh2 in the fine roots increased nitrogen agronomic efficiency. The synergistic expression of the ammonium transporter gene and gdh3 suggests that biochar may be beneficial to amino acid metabolism.

The phosphorylated pathway of serine biosynthesis links plant growth with nitrogen metabolism.

Because it is the precursor for various essential cellular components, the amino acid serine is indispensable for every living organism. In plants, serine is synthesized by two major pathways: photorespiration and the phosphorylated pathway of serine biosynthesis (PPSB). However, the importance of these pathways in providing serine for plant development is not fully understood. In this study, we examine the relative contributions of photorespiration and PPSB to providing serine for growth and metabolism in the C3 model plant Arabidopsis thaliana. Our analyses of cell proliferation and elongation reveal that PPSB-derived serine is indispensable for plant growth and its loss cannot be compensated by photorespiratory serine biosynthesis. Using isotope labeling, we show that PPSB-deficiency impairs the synthesis of proteins and purine nucleotides in plants. Furthermore, deficiency in PPSB-mediated serine biosynthesis leads to a strong accumulation of metabolites related to nitrogen metabolism. This result corroborates 15N-isotope labeling in which we observed an increased enrichment in labeled amino acids in PPSB-deficient plants. Expression studies indicate that elevated ammonium uptake and higher glutamine synthetase/glutamine oxoglutarate aminotransferase (GS/GOGAT) activity causes this phenotype. Metabolic analyses further show that elevated nitrogen assimilation and reduced amino acid turnover into proteins and nucleotides are the most likely driving forces for changes in respiratory metabolism and amino acid catabolism in PPSB-deficient plants. Accordingly, we conclude that even though photorespiration generates high amounts of serine in plants, PPSB-derived serine is more important for plant growth and its deficiency triggers the induction of nitrogen assimilation, most likely as an amino acid starvation response.