Changes In Primary Metabolism Research Articles

Changes in primary metabolism and associated gene expression during host-pathogen interaction in clubroot resistance of Brassica napus.

The role of primary metabolism during Brassica napus-Plasmodiophora brassicae interaction leading to clubroot resistance has not yet been investigated thoroughly. In this study, we investigated some of the primary metabolites and their derivatives as well as expression of the genes involved in their biosynthesis to decipher this host-pathogen interaction. For this, two sets (clubroot resistant and susceptible) of canola lines were inoculated with P. brassicae pathotype 3A to investigate the endogenous levels of primary metabolites at 7-, 14-, and 21-days after inoculation (DAI). The associated pathways were curated, and expression of the selected genes was analyzed using qRT-PCR. Our results suggested the possible involvement of polyamines (spermidine and spermine) in clubroot susceptibility. Some of the amino acids were highly abundant at 7- or 14-DAI in both resistant and susceptible lines; however, glutamine and the amino acid derivative phenylethylamine showed higher endogenous levels in the resistant lines at later stages of infection. Organic acids such as malic, fumaric, succinic, lactic and citric acids were abundant in the susceptible lines. Conversely, the abundance of salicylic acid (SA) and the expression of benzoate/salicylate carboxyl methyltransferase (BSMT) were higher in the resistant lines at the secondary stage of infection. A reduced disease severity index and gall size were observed when exogenous SA (1.0 mM) was applied to susceptible B. napus; this further supported the role of SA in clubroot resistance. In addition, a higher accumulation of fatty acids and significant upregulation of the pathway genes, glycerol-3-phosphate dehydrogenase (GPD) and amino alcohol phosphotransferase (AAPT) were observed in the resistant lines at 14- and 21-DAI. In contrast, some of the fatty acid derivatives such as phosphatidylcholines represented a lower level in the resistant lines. In conclusion, our findings provided additional insights into the possible involvement of primary metabolites and their derivatives in clubroot resistance.

GLUD1 determines murine muscle stem cell fate by controlling mitochondrial glutamate levels.

Muscle stem cells (MuSCs) enable muscle growth and regeneration after exercise or injury, but how metabolism controls their regenerative potential is poorly understood. We describe that primary metabolic changes can determine murine MuSC fate decisions. We found that glutamine anaplerosis into the tricarboxylic acid (TCA) cycle decreases during MuSC differentiation and coincides with decreased expression of the mitochondrial glutamate deaminase GLUD1. Deletion of Glud1 in proliferating MuSCs resulted in precocious differentiation and fusion, combined with loss of self-renewal invitro and invivo. Mechanistically, deleting Glud1 caused mitochondrial glutamate accumulation and inhibited the malate-aspartate shuttle (MAS). The resulting defect in transporting NADH-reducing equivalents into the mitochondria induced compartment-specific NAD+/NADH ratio shifts. MAS activity restoration or directly altering NAD+/NADH ratios normalized myogenesis. In conclusion, GLUD1 prevents deleterious mitochondrial glutamate accumulation and inactivation of the MAS in proliferating MuSCs. It thereby acts as a compartment-specific metabolic brake on MuSC differentiation.

Integrated transcriptomic and metabolomic analyses reveals the molecular bases of alfalfa regrowth processes of new shoots after cutting under different water and nitrogen availability

Alfalfa regrowth is a complex process that is frequently constrained by deficits in water and N fertilisers after cutting. However, the transcriptomic regulatory networks and primary metabolic changes during regrowth are largely unknown. The alfalfa regrowth biomass responses to water deficiency and water deficiency supplied with N fertilisers and corresponding transcriptional and primary metabolic changes in new leaves and roots were systematically explored at 9 and 14 d after cutting. Water deficiency significantly decreased the alfalfa regrowth biomass. Integrated transcriptomic and metabolomic analysis demonstrated that osmoprotection related genes, sucrose and starch metabolic pathways, hormone biosynthesis and signalling pathways, and key transcription factors in new leaves and roots are responsible for alfalfa regrowth under water deficiency. Furthermore, alfalfa regrowth biomass was significantly improved in the presence of N fertilisers under water deficiency through multigene network coordination and metabolic pathway rearrangement. Interestingly, differentially expressed genes expression occurred predominantly in roots at 9 d after cutting, while it mainly occurred in the new leaves at 14 d after cutting. This study revealed the complex molecular mechanisms of alfalfa regrowth and provided theoretical support for optimal irrigation and N fertiliser strategies that will achieve maximum biomass of alfalfa in dryland farming areas.

Combined transcriptomic and pangenomic analyses guide metabolic amelioration to enhance tiancimycins production.

Exploration of high-yield mechanism is important for further titer improvement of valuable antibiotics, but how to achieve this goal is challenging. Tiancimycins (TNMs) are anthraquinone-fused enediynes with promising drug development potentials, but their prospective applications are limited by low titers. This work aimed to explore the intrinsic high-yield mechanism in previously obtained TNMs high-producing strain Streptomyces sp. CB03234-S for the further titer amelioration of TNMs. First, the typical ribosomal RpsL(K43N) mutation in CB03234-S was validated to be merely responsible for the streptomycin resistance but not the titer improvement of TNMs. Subsequently, the combined transcriptomic, pan-genomic and KEGG analyses revealed that the significant changes in the carbon and amino acid metabolisms could reinforce the metabolic fluxes of key CoA precursors, and thus prompted the overproduction of TNMs in CB03234-S. Moreover, fatty acid metabolism was considered to exert adverse effects on the biosynthesis of TNMs by shunting and reducing the accumulation of CoA precursors. Therefore, different combinations of relevant genes were respectively overexpressed in CB03234-S to strengthen fatty acid degradation. The resulting mutants all showed the enhanced production of TNMs. Among them, the overexpression of fadD, a key gene responsible for the first step of fatty acid degradation, achieved the highest 21.7 ± 1.1mg/L TNMs with a 63.2% titer improvement. Our studies suggested that comprehensive bioinformatic analyses are effective to explore metabolic changes and guide rational metabolic reconstitution for further titer improvement of target products. KEY POINTS: • Comprehensive bioinformatic analyses effectively reveal primary metabolic changes. • Primary metabolic changes cause precursor enrichment to enhance TNMs production. • Strengthening of fatty acid degradation further improves the titer of TNMs.

Nitrogen deficiency stimulates cannabinoid biosynthesis in medical cannabis plants by inducing a metabolic shift towards production of low-N metabolites

The primary and secondary metabolism of plants is closely connected to the resources supplied and obtained by the plant, including their mineral nutrition. We recently reported that nitrogen (N) deficiency enhances the production of terpenoids and cannabinoids, the unique biologically-active secondary metabolites in medical cannabis plants. Knowledge-gaps concerning effects of N supply on primary metabolism in cannabis hinder understanding of the interrelations between N inputs and biosynthesis of the therapeutic secondary metabolites. The present study therefore evaluated the hypothesis that a decrease in the plant’s C (carbon): N ratio by N-limitation, induces a shift in the plant metabolism towards lower production of N-containing metabolites and higher production of metabolites that do not contain N. Effects of N input levels (30, 80, 160, 240, and 320 mg L−1) on total C and N concentrations and C: N ratio in leaves and inflorescences of medical cannabis plants were studied; and gas chromatography-mass spectrometry (GC-MS) analysis of primary metabolites added to the analyses of secondary metabolites. Elevation of N supply resulted in an increase in total N and N-containing compounds (chlorophylls and most amino acids), and decreased total C and compounds that do not contain N, such as sugars (fructose, glucose, and xylose), and phosphates (phosphate and glucose-6-phosphate) in both inflorescences and leaves. In the inflorescences, the elevation of N input also decreased total cannabinoids, phenols, and flavonoids, that do not contain N. Integrating the metabolic datasets revealed positive correlations between C sources (fructose and glucose) and most of the cannabinoids and terpenoids; the latter were negatively correlated with N-compounds (most amino acids). Taken together, these results suggest that elevated N supply induce a metabolic shift in the inflorescences towards increased production of N-compounds via deflecting the C sources from the biologically active compounds. In addition, the cannabis leaf was found to be more sensitive than the inflorescence to N supply, presenting greater changes in primary metabolism and more coordinated metabolic associations. These findings highlight the importance of adequate and precise N nutrition for standardization of the therapeutic-metabolite profile and for preventing undesirable metabolic repartitioning in medical cannabis plants.

Postharvest UV-B exposure drives changes in primary metabolism, phenolic concentration, and volatilome profile in berries of different grape (Vitis vinifera L.) varieties.

The ultraviolet-B (UV-B) radiation can alter grape metabolism during berry development, but little is known on the effect of postharvest UV-B exposure. In this study, we evaluated the effect of postharvest UV-B exposure on berry primary and secondary metabolites in four grapevine varieties (Aleatico, Moscato bianco, Sangiovese, and Vermentino) in order to evaluate the possibility to increase the grape quality and its nutraceutical properties. The treatment did not significantly affect the berry primary metabolism in terms of organic acids, carbohydrates, and amino acids profile, regardless of the variety. UV-B exposure reduced the total anthocyanin content, particularly the tri-substituted and di-substituted forms in Aleatico and Sangiovese, respectively. An overall negative effect of UV-B irradiation on the flavonols profile of Aleatico, Moscato bianco, and Vermentino berries was found, whereas it enhanced the quercetin, myricetin and kaempferol concentration in Sangiovese. The free fraction of berry volatile organic compounds increased in UV-B-treated Aleatico and Moscato bianco berries, especially C13 -norisoprenoids and volatile phenols, as well as key monoterpenes, such as the linalool derivatives. However, higher concentrations of glycosylated monoterpenes and C13 -norisoprenoids were measured in Sangiovese and Vermentino berries treated with UV-B. This study provides new insights on the effect of postharvest UV-B radiation on berry secondary metabolism, highlighting a different modulation between varieties and suggesting the potential use of this technique to increase some nutraceutical and quality characteristics of grape berry. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Temporal segregation of biosynthetic processes is responsible for metabolic oscillations during the budding yeast cell cycle

Many cell biological and biochemical mechanisms controlling the fundamental process of eukaryotic cell division have been identified; however, the temporal dynamics of biosynthetic processes during the cell division cycle are still elusive. Here, we show that key biosynthetic processes are temporally segregated along the cell cycle. Using budding yeast as a model and single-cell methods to dynamically measure metabolic activity, we observe two peaks in protein synthesis, in the G1 and S/G2/M phase, whereas lipid and polysaccharide synthesis peaks only once, during the S/G2/M phase. Integrating the inferred biosynthetic rates into a thermodynamic-stoichiometric metabolic model, we find that this temporal segregation in biosynthetic processes causes flux changes in primary metabolism, with an acceleration of glucose-uptake flux in G1 and phase-shifted oscillations of oxygen and carbon dioxide exchanges. Through experimental validation of the model predictions, we demonstrate that primary metabolism oscillates with cell-cycle periodicity to satisfy the changing demands of biosynthetic processes exhibiting unexpected dynamics during the cell cycle.

Reduced auxin signalling through the cyclophilin gene DIAGEOTROPICA impacts tomato fruit development and metabolism during ripening.

Auxin is an important hormone playing crucial roles during fruit growth and ripening; however, the metabolic impact of changes in auxin signalling during tomato (Solanum lycopersicum L.) ripening remains unclear. Here, we investigated the significance of changes in auxin signalling during different stages of fruit development by analysing changes in tomato fruit quality and primary metabolism using mutants with either lower or higher auxin sensitivity [diageotropica (dgt) and entire mutants, respectively]. Altered auxin sensitivity modifies metabolism, through direct impacts on fruit respiration and fruit growth. We verified that the dgt mutant plants exhibit reductions in fruit set, total fruit dry weight, fruit size, number of seeds per fruit, and fresh weight loss during post-harvest. Sugar accumulation was associated with delayed fruit ripening in dgt, probably connected with reduced ethylene levels and respiration, coupled with a lower rate of starch degradation. In contrast, despite exhibiting parthenocarpy, increased auxin perception (entire) did not alter fruit ripening, leading to only minor changes in primary metabolism. By performing a comprehensive analysis, our results connect auxin signalling and metabolic changes during tomato fruit development, indicating that reduced auxin signalling led to extensive changes in sugar concentration and starch metabolism during tomato fruit ripening.

Crosstalk between Selenium and Sulfur Is Associated with Changes in Primary Metabolism in Lettuce Plants Grown under Se and S Enrichment.

This study investigated the beneficial effects of selenium (Se) and sulfur (S) enrichment on the primary metabolism in butterhead lettuce. The plants were treated with three levels of Se via foliar application in the presence of two S levels in the nutrient solution under greenhouse conditions. The lettuce plants that were exposed to the lower selenate level (1.3 μM) in combination with the adequate and high S supplies (1 and 2 mM, respectively) accumulated 38.25 ± 0.38 µg Se g−1 DM and 47.98 ± 0.68 µg Se g−1 DM, respectively. However, a dramatic increase in the Se concentration (122.38 ± 5.07 µg Se g−1 DM, and 146.71 ± 5.43 µg Se g−1 DM, respectively) was observed in the lettuce heads that were exposed to the higher selenate foliar application (3.8 μM) in response to the varied sulfate concentrations (S1 and S2, respectively). Under higher Se and S supplies in the lettuce plants, the levels of organic acids, including malic acid and citric acid, decreased therein to 25.7 ± 0.5 and 3.9 ± 0.3 mg g−1 DM, respectively, whereas, in the plants that were subjected to adequate S and lower Se fertilization, the malic acid, and citric acid levels significantly increased to 47.3 ± 0.4 and 11.8 ± 0.4 mg g−1 DM, respectively. The two Se levels (1.3 and 3.8 μM) under the S1 conditions also showed higher concentrations of water-soluble sugars, including glucose and fructose (70.8.4 ± 1.1 and 115.0 ± 2.1 mg g−1 DM; and 109.4 ± 2.1 and 161.1 ± 1.0 mg g−1 DM, respectively), compared to the control. As with the glucose and fructose, the amino acids (Asn, Glu, and Gln) exhibited strikingly higher levels (48.7 ± 1.1 μmol g−1 DM) under higher S and Se conditions. The results presented in this report reveal that the “crosstalk” between Se and S exhibited a unique synergistic effect on the responses to the amino acids and the soluble sugar biosynthesis under Se and S enrichment. Additionally, the Se-and-S crosstalk could have an important implication on the final nutritional value and quality of lettuce plants.

Exploratory Growth in Streptomyces venezuelae Involves a Unique Transcriptional Program, Enhanced Oxidative Stress Response, and Profound Acceleration in Response to Glycerol.

Exploration is a recently discovered mode of growth and behavior exhibited by some Streptomyces species that is distinct from their classical sporulating life cycle. While much has been uncovered regarding initiating environmental conditions and phenotypic outcomes of exploratory growth, how this process is coordinated at a genetic level remains unclear. We used RNA sequencing to survey global changes in the transcriptional profile of exploring cultures over time in the model organism Streptomyces venezuelae. Transcriptomic analyses revealed widespread changes in gene expression impacting diverse cellular functions. Investigations into differentially expressed regulatory elements revealed specific groups of regulatory factors to be impacted, including the expression of several extracytoplasmic function (ECF) sigma factors, second messenger signaling pathways, and members of the whiB-like (wbl) family of transcription factors. Dramatic changes were observed among primary metabolic pathways, especially among respiration-associated genes and the oxidative stress response; enzyme assays confirmed that exploring cultures exhibit an enhanced oxidative stress response compared with classically growing cultures. Changes in the expression of the glycerol catabolic genes in S. venezuelae led to the discovery that glycerol supplementation of the growth medium promotes a dramatic acceleration of exploration. This effect appears to be unique to glycerol as an alternative carbon source, and this response is broadly conserved across other exploration-competent species. IMPORTANCE Exploration represents an alternative growth strategy for Streptomyces bacteria and is initiated in response to other microbes or specific environmental conditions. Here, we show that entry into exploration involves comprehensive transcriptional reprogramming, with an emphasis on changes in primary metabolism and regulatory/signaling functions. Intriguingly, a number of transcription factor classes were downregulated upon entry into exploration. In contrast, respiration-associated genes were strongly induced, and this was accompanied by an enhanced oxidative stress response. Notably, our transcriptional analyses suggested that glycerol may play a role in exploration, and we found that glycerol supplementation dramatically enhanced the exploration response in many streptomycetes. This work sheds new light on the regulatory and metabolic cues that influence a fascinating new microbial behavior.

Cell death and changes in primary metabolism: the onset of defence in Eucalyptus in the war against Leptocybe invasa.

Here, we investigated changes in primary metabolism and cell death around oviposition sites in two hybrid clones of Eucalyptus with different degrees of resistance to Leptocybe invasa Fisher & La Salle (Hymenoptera: Eulophidae), as well as tolerance to water deficiency. We showed that apices of the resistant clone with oviposition had a higher content of amino acids, organic acids and the compound putrescine compared with those of the susceptible clone with oviposition. By contrast, apices of the resistant clone with oviposition had lower sugar and pyruvate organic acid content than those of the susceptible clone with oviposition. Small areas of necrosis were induced around the oviposition sites in the stem apices of Eucalyptus 24 h after infestation. The resistant clone developed larger necrotic areas that showed progressive increases 24-72 h after infestation compared with the susceptible clone, in which cell death was significantly lower and no changes were observed in necrotic area over time. Thus, the programmed death of cells around the egg, modulated by several amino acids, is likely the first defence response of Eucalyptus against L. invasa. Our results serve as the basis for the early identification of key metabolites produced in plants in defence against galling insects. © 2022 Society of Chemical Industry.

Arbuscular Mycorrhizal Symbiosis Triggers Major Changes in Primary Metabolism Together With Modification of Defense Responses and Signaling in Both Roots and Leaves of Vitis vinifera.

Grapevine (Vitis vinifera L.) is one of the most important crops worldwide but is subjected to multiple biotic and abiotic stresses, especially related to climate change. In this context, the grapevine culture could take advantage of symbiosis through association with arbuscular mycorrhizal fungi (AMF), which are able to establish symbiosis with most terrestrial plants. Indeed, it is well established that mycorrhization improves grapevine nutrition and resistance to stresses, especially water stress and resistance to root pathogens. Thus, it appears essential to understand the effect of mycorrhization on grapevine metabolism and defense responses. In this study, we combined a non-targeted metabolomic approach and a targeted transcriptomic study to analyze changes induced in both the roots and leaves of V. vinifera cv. Gewurztraminer by colonization with Rhizophagus irregularis (Ri). We showed that colonization of grapevine with AMF triggers major reprogramming of primary metabolism in the roots, especially sugar and fatty acid metabolism. On the other hand, mycorrhizal roots had decreased contents of most sugars and sugar acids. A significant increase in several fatty acids (C16:1, linoleic and linolenic acids and the C20 arachidonic and eicosapentaenoic acids) was also detected. However, a downregulation of the JA biosynthesis pathway was evidenced. We also found strong induction of the expression of PR proteins from the proteinase inhibitor (PR6) and subtilase (PR7) families in roots, suggesting that these proteins are involved in the mycorrhiza development but could also confer higher resistance to root pathogens. Metabolic changes induced by mycorrhization were less marked in leaves but involved higher levels of linoleic and linolenic acids and decreased sucrose, quinic, and shikimic acid contents. In addition, Ri colonization resulted in enhanced JA and SA levels in leaves. Overall, this study provides a detailed picture of metabolic changes induced by AMF colonization in a woody, economically important species. Moreover, stimulation of fatty acid biosynthesis and PR protein expression in roots and enhanced defense hormone contents in leaves establish first insight in favor of better resistance of grapevine to various pathogens provided by AMF colonization.

Specific leaf area is modulated by nitrogen via changes in primary metabolism and parenchymal thickness in pepper.

Nitrogen promotes changes in SLA through metabolism and anatomical traits in Capsicum plants. Specific leaf area (SLA) is a key trait influencing light interception and light use efficiency that often impacts plant growth and production. SLA is a key trait explaining growth variations of plant species under different environments. Both light and nitrogen (N) supply are important determinants of SLA. To better understand the effect of irradiance level and N on SLA in Capsicum chinense, we evaluated primary metabolites and morphological traits of two commercial cultivars (Biquinho and Habanero) in response to changes in both parameters. Both genotypes showed increased SLA with shading, and a decrease in SLA in response to increased N supply, however, with Habanero showing a stable SLA in the range of N deficiency to sufficient N doses. Correlation analyses indicated that decreased SLA in response to higher N supply was mediated by altered amino acids, protein, and starch levels, influencing leaf density. Moreover, in the range of moderate N deficiency to N sufficiency, both genotypes exhibited differences in SLA response, with Biquinho and Habanero displaying alterations on palisade and spongy parenchyma, respectively. Altogether, the results suggest that SLA responses to N supply are modulated by the balance between certain metabolites content and genotype-dependent changes in the parenchyma cells influencing leaf thickness and density.

Dynamic Inventory of Intermediate Metabolites of Cyanobacteria in a Diurnal Cycle.

SummaryCyanobacteria are gaining importance both as hosts for photoautotrophic production of chemicals and as model systems for studies of diurnal lifestyle. The proteome and transcriptome of cyanobacteria have been closely examined under diurnal growth, whereas the downstream effects on the intermediary metabolism have not received sufficient attention. The present study focuses on identifying the cellular metabolites whose inventories undergo dramatic changes in a fast-growing cyanobacterium, Synechococcus elongatus PCC 11801. We identified and quantified 67 polar metabolites, whose inventory changes significantly during diurnal growth, with some metabolites changing by 100-fold. The Calvin-Benson-Bassham cycle intermediates peak at midday to support fast growth. The hitherto unexplored γ-glutamyl peptides act as reservoirs of amino acids. Interestingly, several storage molecules or their precursors accumulate during the dark phase, dispelling the notion that all biosynthetic activity takes place in the light phase. Our results will guide metabolic modeling and strain engineering of cyanobacteria.

Redox Homeostasis and Signaling in a Higher-CO2 World.

Rising CO2 concentrations and their effects on plant productivity present challenging issues. Effects on the photosynthesis/photorespiration balance and changes in primary metabolism are known, caused by the competitive interaction of CO2 and O2 at the active site of ribulose-1,5-bisphosphate carboxylase/oxygenase. However, impacts on stress resistance are less clear. Reactive oxygen species are key players in biotic and abiotic stress responses, but there is no consensus on whether elevated CO2 constitutes a stress. Although high CO2 increases yield in C3 plants, it can also increase cellular oxidation and activate phytohormone defense pathways. Reduction-oxidation processes play key roles in acclimation to high CO2, with specific enzymes acting in compartment-specific signaling. Traditionally, acclimation to high CO2 has been considered in terms of altered carbon gain, but emerging evidence suggests that CO2 is a signal as well as a substrate. Some CO2 effects on defense are likely mediated independently of primary metabolism. Nonetheless, primary photosynthetic metabolism is highly integrated with defense and stress signaling pathways, meaning that plants will be able to acclimate to the changing environment over the coming decades.

Downregulation of a Mitochondrial NAD+ Transporter (NDT2) Alters Seed Production and Germination in Arabidopsis.

Despite the fundamental importance of nicotinamide adenine dinucleotide (NAD+) for metabolism, the physiological roles of NAD+ carriers in plants remain unclear. We previously characterized the Arabidopsis thaliana gene (At1g25380), named AtNDT2, encoding a protein located in the mitochondrial inner membrane, which imports NAD+ from the cytosol using ADP and AMP as counter-exchange substrates for NAD+. Here, we further investigated the physiological roles of NDT2, by isolating a T-DNA insertion line, generating an antisense line and characterizing these genotypes in detail. Reduced NDT2 expression affected reproductive phase by reducing total seed yield. In addition, reduced seed germination and retardation in seedling establishment were observed in the mutant lines. Moreover, remarkable changes in primary metabolism were observed in dry and germinated seeds and an increase in fatty acid levels was verified during seedling establishment. Furthermore, flowers and seedlings of NDT2 mutants displayed upregulation of de novo and salvage pathway genes encoding NAD+ biosynthesis enzymes, demonstrating the transcriptional control mediated by NDT2 activity over these genes. Taken together, our results suggest that NDT2 expression is fundamental for maintaining NAD+ balance amongst organelles that modulate metabolism, physiology and developmental processes of heterotrophic tissues.

GABA negatively regulates adventitious root development in poplar.

γ-Aminobutyric acid (GABA) influences plant growth, but little is known about how this metabolite regulates adventitious root (AR) development. Here, we investigate the effects of GABA on ARs using poplar lines overexpressing glutamate decarboxilase 2 (GAD2) and by treating poplar stem cuttings with exogenous GABA or vigabatrin (VGB; a specific GABA transaminase inhibitor). Endogenous GABA accumulation not only inhibited AR growth, but it also suppressed or delayed AR formation. Anatomical observations revealed that the GABA and VGB treatments resulted in a 1 d delay in the formation of AR primordia and the appearance of ARs. This delay coincided with changes in primary metabolism, including transient increases in hexose and amino acid levels. GABA-dependent changes in the expression of genes related to hormone synthesis and signalling, as well as analysis of hormone levels revealed that ethylene-dependent pathways were decreased at the earliest stage of AR formation. In contrast, auxin and abscisic acid were increased at 1-5 d as well as GA4 over a 5 d period of AR formation. These results demonstrate that GABA plays a crucial role in AR development. Evidence is presented demonstrating that GABA can interact with hormone-related pathways as well as carbon/nitrogen metabolism. These findings also elucidate the functions of GABA in plant development.

Ammonium mediated changes in carbon and nitrogen metabolisms induce resistance against Pseudomonas syringae in tomato plants

Predominant NH4+ nutrition causes an “ammonium syndrome” that induces metabolic changes and thereby provides resistance against Pseudomonas syringae infection through the activation of systemic acquired acclimation (SAA). Hence, to elucidate the mechanisms underlying NH4+-mediated SAA, the changes in nutrient balance and C and N skeletons were studied in NH4+-treated plants upon infection by P. syringae. A general decrease in cation and an increase in anion levels was observed in roots and leaves of NH4+-treated plants. Upon NH4+-based nutrition and infection, tomato leaves showed an accumulation of S, P, Zn, and of Mn. Mn accumulation might be required for ROS detoxification since it acts as a cofactor of superoxide dismutase (SOD). Primary metabolism was modified in both tissues of NH4+-fed plants to counteract NH4+ toxicity by decreasing TCA intermediates. A significant increase in Arg, Gln, Asn, Lys, Tyr, His and Leu was observed in leaves of NH4+-treated plants. The high level of the putrescine precursor Arg hints towards the importance of the Glu pathway as a key metabolic check-point in NH4+-treated and infected plants. Taken together, NH4+-fed plants displayed a high level of basal responses allowing them to activate SAA and to trigger defense responses against P. syringae through nutrient imbalances and changes in primary metabolism.

Tecia solanivora infestation increases tuber starch accumulation in Pastusa Suprema potatoes.

In response to infestation with larvae of the Guatemalan tuber moth (Tecia solanivora), some Solanum tuberosum (potato) varieties exhibit an overcompensation response, whereby the total dry mass of uninfested tubers is increased. Here, we describe early responses, within the first few days, of T. solanivora feeding, in the Colombian potato variety Pastusa Suprema. Non-targeted metabolite profiling showed significant secondary metabolism changes in T. solanivora-infested tubers, but not in uninfested systemic tubers. In contrast, changes in primary metabolism were greater in uninfested systemic tubers than in the infested tubers, with a notable 80% decline in systemic tuber sucrose levels within 1 d of T. solanivora infestation. This suggested either decreased sucrose transport from the leaves or increased sink strength, i.e., more rapid sucrose to starch conversion in the tubers. Increased sucrose synthesis was indicated by higher rubisco activase and lower starch synthase gene expression in the leaves of infested plants. Elevated sink strength was demonstrated by 45% more total starch deposition in systemic tubers of T. solanivora-infested plants compared to uninfested control plants. Thus, rather than investing in increased defense of uninfested tubers, Pastusa Suprema promotes deposition of photoassimilates in the form of starch as a response to T. solanivora infestation.

1H-NMR metabolomic profiling reveals a distinct metabolic recovery response in shoots and roots of temporarily drought-stressed sugar beets.

Yield formation in regions with intermittent drought periods depends on the plant’s ability to recover after cessation of the stress. The present work assessed differences in metabolic recovery of leaves and roots of drought-stressed sugar beets with high temporal resolution. Plants were subjected to drought for 13 days, and rewatered for 12 days. At one to two-day intervals, plant material was harvested for untargeted 1H-NMR metabolomic profiling, targeted analyses of hexose-phosphates, starch, amino acids, nitrate and proteins, and physiological measurements including relative water content, osmotic potential, electrolyte leakage and malondialdehyde concentrations. Drought triggered changes in primary metabolism, especially increases in amino acids in both organs, but leaves and roots responded with different dynamics to rewatering. After a transient normalization of most metabolites within 8 days, a second accumulation of amino acids in leaves might indicate a stress imprint beneficial in upcoming drought events. Repair mechanisms seemed important during initial recovery and occurred at the expense of growth for at least 12 days. These results indicate that organ specific metabolic recovery responses might be related to distinct functions and concomitant disparate stress levels in above- and belowground organs. With respect to metabolism, recovery was not simply a reversal of the stress responses.