MicroRNA169 integrates multiple factors to modulate plant growth and abiotic stress responses.

MicroRNA169 (miR169) has been implicated in multi-stress regulation in annual species such as Arabidopsis, maize and rice. However, there is a lack of experimental functional and mechanistic studies of miR169 in plants, especially in perennial species, and its impact on plant growth and development remains unexplored. Creeping bentgrass (Agrostis stolonifera L.) is a C3 cool-season perennial turfgrass of significant environmental and economic importance. In this study, we generated both miR169 overexpression and knockdown transgenic creeping bentgrass lines. We found that miR169 acts as a positive regulator in abiotic stress responses but is negatively associated with plant growth and development, playing multiple critical roles in the growth and environmental adaptation of creeping bentgrass. These roles include differentiated spatial hormone accumulation patterns associated with growth and stress accommodation, elevated antioxidant activity that alleviates oxidative damage induced by stress, ion-channelling membrane components for maintaining homeostasis under saline conditions, and potential cross-talks with stress-regulating transcription factors such as AsHsfA and AsWRKYs. Our results unravel the role of miR169 in modulating plant development and stress responses in perennial grass species. This underlines the potential of manipulating miR169 to generate crop cultivars with desirable traits to meet diverse agricultural demands.

Molecular-level carbon traits underlie the multidimensional fine root economics space.

Carbon influences the evolution and functioning of plants and their roots. Previous work examining a small number of commonly measured root traits has revealed a global multidimensionality of the resource economics traits in fine roots considering carbon as primary currency but without considering the diversity of carbon-related traits. To address this knowledge gap, we use data from 66 tree species from a tropical forest to illustrate that root economics space co-varies with a novel molecular-level traits space based on nuclear magnetic resonance. Thinner fine roots exhibit higher proportions of carbohydrates and lower diversity of molecular carbon than thicker roots. Mass-denser fine roots have more lignin and aromatic carbon compounds but less bioactive carbon compounds than lighter roots. Thus, the transition from thin to thick fine roots implies a shift in the root carbon economy from 'do-it-yourself' soil exploration to collaboration with mycorrhizal fungi, while the shift from light to dense fine roots emphasizes a shift from acquisitive to conservative root strategy. We reveal a previously undocumented role of molecular-level carbon traits that potentially undergird the multidimensional root economics space. This finding offers new molecular insight into the diversity of root form and function, which is fundamental to our understanding of plant evolution, species coexistence and adaptations to heterogeneous environments.

Increased Perfluorooctanesulfonate (PFOS) Toxicity and Accumulation Is Associated with Perturbed Prostaglandin Metabolism and Increased Organic Anion Transport Protein (OATP) Expression.

Perfluorooctanesulfonate (PFOS) is a widespread environmental pollutant with a long half-life and clearly negative outcomes on metabolic diseases such as fatty liver disease and diabetes. Male and female Cyp2b-null and humanized CYP2B6-transgenic (hCYP2B6-Tg) mice were treated with 0, 1, or 10 mg/kg/day PFOS for 21 days, and surprisingly it was found that PFOS was retained at greater concentrations in the serum and liver of hCYP2B6-Tg mice than those of Cyp2b-null mice, with greater differences in the females. Thus, Cyp2b-null and hCYP2B6-Tg mice provide new models for investigating individual mechanisms for PFOS bioaccumulation and toxicity. Overt toxicity was greater in hCYP2B6-Tg mice (especially females) as measured by mortality; however, steatosis occurred more readily in Cyp2b-null mice despite the lower PFOS liver concentrations. Targeted lipidomics and transcriptomics from PFOS-treated Cyp2b-null and hCYP2B6-Tg mouse livers were performed and compared to PFOS retention and serum markers of toxicity using PCA. Several oxylipins, including prostaglandins, thromboxanes, and docosahexaenoic acid metabolites, are associated or inversely associated with PFOS toxicity. Both lipidomics and transcriptomics indicate PFOS toxicity is associated with PPAR activity in all models. GO terms associated with reduced steatosis were sexually dimorphic with lipid metabolism and transport increased in females and circadian rhythm associated genes increased in males. However, we cannot rule out that steatosis was initially protective from PFOS toxicity. Moreover, several transporters are associated with increased retention, probably due to increased uptake. The strongest associations are the organic anion transport proteins (Oatp1a4-6) genes and a long-chain fatty acid transport protein (fatp1), enriched in female hCYP2B6-Tg mice. PFOS uptake was also reduced in cultured murine hepatocytes by OATP inhibitors. The role of OATP1A6 and FATP1 in PFOS transport has not been tested. In summary, Cyp2b-null and hCYP2B6-Tg mice provided unique models for estimating the importance of novel mechanisms in PFOS retention and toxicity.

Binding of Pentagalloyl Glucose to Aortic Wall Proteins: Insights from Peptide Mapping and Simulated Docking Studies.

Pentagalloyl glucose (PGG) is currently being investigated as a non-surgical treatment for abdominal aortic aneurysms (AAAs); however, the molecular mechanisms of action of PGG on the AAA matrix components and the intra-luminal thrombus (ILT) still need to be better understood. To assess these interactions, we utilized peptide fingerprinting and molecular docking simulations to predict the binding of PGG to vascular proteins in normal and aneurysmal aorta, including matrix metalloproteinases (MMPs), cytokines, and fibrin. We performed PGG diffusion studies in pure fibrin gels and human ILT samples. PGG was predicted to bind with high affinity to most vascular proteins, the active sites of MMPs, and several cytokines known to be present in AAAs. Finally, despite potential binding to fibrin, PGG was shown to diffuse readily through thrombus at physiologic pressures. In conclusion, PGG can bind to all the normal and aneurysmal aorta protein components with high affinity, potentially protecting the tissue from degradation and exerting anti-inflammatory activities. Diffusion studies showed that thrombus presence in AAAs is not a barrier to endovascular treatment. Together, these results provide a deeper understanding of the clinical potential of PGG as a non-surgical treatment of AAAs.

Global Metabolome of Palmer Amaranth (Amaranthus palmeri) Populations Highlights the Specificity and Inducibility of Phytochemical Responses to Abiotic Stress.

Commonalities in adaptive responses to abiotic stressors could contribute to the development of cross-resistance in weeds. The degree to which herbicide-induced changes in weeds parallel those induced by other abiotic stress remains unknown. We investigated the specificity of metabolic perturbations induced by glyphosate and drought across three glyphosate-resistant (GR) and two glyphosate-susceptible (GS) biotypes of Palmer amaranth (Amaranthus palmeri) using global metabolomics approaches. Compared to GS-biotypes, in the absence of stress, the GR-biotypes had a higher abundance of primary metabolites, including sugars, nonaromatic amino acids, and organic acids. However, despite having a higher 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene copy number that could upregulate the phenylpropanoid metabolism, the nonstressed GR-biotypes were less abundant in specialized (secondary) metabolites. Under glyphosate stress, 80% of metabolites, including shikimate, that accumulated in GS-biotypes also increased in the GR-biotypes. However, glyphosate triggered the preferential accumulation of glycosides of dihydroxylated and methoxylated flavanols with higher antioxidant potential, and ferulic acid derivatives, specifically in GR-biotypes. The disruption of the shikimate pathway and the accumulation of phenylpropanoids upon glyphosate exposure suggest that the stress response of GR-biotypes could be partly induced. This differential response was less evident in other phytochemical classes and under drought, highlighting that the phytochemical responses are stress-specific rather than biotype-specific.

Comparative uptake, translocation, and plant mediated transport of Tc-99, Cs-133, Np-237, and U-238 in Savannah River Site soil columns for the grass species Andropogon virginicus.

This study examines the ability of the grass species Andropogon virginicus to alter the subsurface transport and redistribution of a suite of radionuclides (99Tc, 133Cs (stable analog for 135Cs and 137Cs), 237Np, 238U) with varying chemical behaviors in a Savannah River Site soil via the use of vegetated and unvegetated soil columns. After an acclimation period, a small volume of solution containing all radionuclides was introduced into the columns via Rhizon© pore water sampling tubes. Plants were grown for an additional 4 weeks before shoots were harvested, and columns were prepared for sampling. Plant presence led to decreased radionuclide release from the columns, mainly due to radionuclide specific combinations of system hydrology differences resulting from plant transpiration as well as plant uptake. For the most mobile radionuclides, 99Tc followed by 237Np, plant presence resulted in significantly different soil concentration profiles between vegetated and unvegetated columns, including notable upward migration for 237Np in columns with plants. Additionally, plant uptake of 99Tc was the greatest of all the radionuclides, with plant tissues containing an average of 44 % of the 99Tc, while plant uptake only accounted for <2 % of 237Np and <0.5 % of 133Cs and 238U in the system. Although overall plant uptake of 133Cs and 238U were similar, the majority of 133Cs taken up by plants was associated with 133Cs already available in the aqueous phase while 238U uptake was mainly associated with the solid phase, meaning that plant activity resulted in a fraction of the native 238U being mobilized and thus, made available for plant uptake. Overall, this study quantified the influence of several plant-mediated physical and biogeochemical factors that have significant influence on radionuclide mobility and transport in this complex system which can be further utilized in future system or site-specific environmental transport and risk assessment models.

The Influence of Iron and Ligand Type on Plutonium Uptake in Two Strains of Hydroponically Grown Corn ( Zea Mays ).

ABSTRACT: This work investigates the uptake and root-shoot transport of plutonium (Pu) and iron (Fe) in corn ( Zea mays ) to gain insight into the Pu uptake pathway. Plutonium has no known biological function in plants yet may feasibly enter plants through the uptake pathway used by Fe (an essential nutrient), as these two elements have similar chemical properties. A series of experiments was conducted in which two hydroponically grown corn strains (one normal and one deficient in the transporter protein for Fe) were exposed to varying concentrations of complexed Pu and Fe. Results suggest that while Fe did inhibit Pu uptake to a certain extent, Pu was able to use alternative uptake pathways. In a 10 ppb Pu:1 ppb Fe hydroponic solution, all shoots had detectable shoot Pu concentrations compared to only 22% of plants when the Fe concentration was raised to 10 ppb. While root Pu accumulation was reduced for the corn strain deficient in the Fe transporter protein at lower Pu media concentrations, there were no differences at higher Pu concentrations, signifying the existence of substitute transport routes. A comparison of citrate and deferoxamine B (DFOB) ligand influence found that Pu complexed with DFOB remained in the roots of the plant, while movement of Pu into the shoots of the plant was more prevalent with the Pu-citrate complex. This study advances understanding of the behavior and mobility of Pu in the terrestrial environment and specifically the interactions between Pu and an essential nutrient in a common crop species.

Characterizing natural variability of lignin abundance and composition in fine roots across temperate trees: a comparison of analytical methods.

Lignin is an important root chemical component that is widely used in biogeochemical models to predict root decomposition. Across ecological studies, lignin abundance has been characterized using both proximate and lignin-specific methods, without much understanding of their comparability. This uncertainty in estimating lignin limits our ability to comprehend the mechanisms regulating root decomposition and to integrate lignin data for large-scale syntheses. We compared five methods of estimating lignin abundance and composition in fine roots across 34 phylogenetically diverse tree species. We also assessed the feasibility of high-throughput techniques for fast-screening of root lignin. Although acid-insoluble fraction (AIF) has been used to infer root lignin and decomposition, AIF-defined lignin content was disconnected from the lignin abundance estimated by techniques that specifically measure lignin-derived monomers. While lignin-specific techniques indicated lignin contents of 2-10% (w/w) in roots, AIF-defined lignin contents were c. 5-10-fold higher, and their interspecific variation was found to be largely unrelated to that determined using lignin-specific techniques. High-throughput pyrolysis-gas chromatography-mass spectrometry, when combined with quantitative modeling, accurately predicted lignin abundance and composition, highlighting its feasibility for quicker assessment of lignin in roots. We demonstrate that AIF should be interpreted separately from lignin in fine roots as its abundance is unrelated to that of lignin polymers. This study provides the basis for informed decision-making with respect to lignin methodology in ecology.

Cross-inoculation of rhizobiome from a congeneric ruderal plant imparts drought tolerance in maize (Zea mays) through changes in root morphology and proteome.

Rhizobiome confer stress tolerance to ruderal plants, yet their ability to alleviate stress in crops is widely debated, and the associated mechanisms are poorly understood. We monitored the drought tolerance of maize (Zea mays) as influenced by the cross-inoculation of rhizobiota from a congeneric ruderal grass Andropogon virginicus (andropogon-inoculum), and rhizobiota from organic farm maintained under mesic condition (organic-inoculum). Across drought treatments (40% field capacity), maize that received andropogon-inoculum produced two-fold greater biomass. This drought tolerance translated to a similar leaf metabolomic composition as that of the well-watered control (80% field capacity) and reduced oxidative damage, despite a lower activity of antioxidant enzymes. At a morphological-level, drought tolerance was associated with an increase in specific root length and surface area facilitated by the homeostasis of phytohormones promoting root branching. At a proteome-level, the drought tolerance was associated with upregulation of proteins related to glutathione metabolism and endoplasmic reticulum-associated degradation process. Fungal taxa belonging to Ascomycota, Mortierellomycota, Archaeorhizomycetes, Dothideomycetes, and Agaricomycetes in andropogon-inoculum were identified as potential indicators of drought tolerance. Our study provides a mechanistic understanding of the rhizobiome-facilitated drought tolerance and demonstrates a better path to utilize plant-rhizobiome associations to enhance drought tolerance in crops.

Elevational divergence in pigmentation plasticity is associated with selection and pigment biochemistry.

Phenotypic plasticity is predicted to evolve in environmentally variable habitats, or those experiencing a high frequency of strong selection. The evolution of plasticity may however be constrained by costs or physiological limitations. In flowers, UV-absorbing pigmentation ameliorates UV damage to pollen, and is linked with elevated UV exposure. Whether plasticity contributes to this pattern remains unclear. Petals of Argentina anserina have larger UV-absorbing petal areas at high elevations where they experience higher and more variable UV exposure than low elevations. We measured UV-induced pigmentation plasticity in high- and low-elevation populations (hereafter, "high," "low"), and selection on pigmentation via male fitness. We dissected UV pigment biochemistry using metabolomics to explore biochemical mechanisms underlying plasticity. High displayed positive UV-induced pigmentation plasticity but low lacked plasticity. Selection favored elevated pigmentation under UV in high, supporting adaptive plasticity. In high, UV absorption was conferred by flavonoids produced in one flavonoid pathway branch. However, in low, UV absorption was associated with many compounds spanning multiple branches. Elevated plasticity was thus associated with reduced pigment diversity. These results are consistent with adaptive floral pigmentation plasticity in more extreme and variable environments. We discuss how biochemical underpinnings of pigmentation may permit or constrain the evolution of pigmentation plasticity.

Persistence and metabolism of the diamide insecticide cyantraniliprole in tomato plants.

Plant uptake and metabolism of pesticides are complex and dynamic processes, which contribute to the overall toxicity of the pesticides. We investigated the metabolic fate of cyantraniliprole, a new diamide class of insecticide, during various growth stages of tomato. Cyantraniliprole was the major residue in leaves, flowers, and fruits, with the relative metabolite-to-parent ratios maintained at < 10% up to 28 days after treatment (DAT). Mature leaves contained consistently higher residues of cyantraniliprole than young leaves throughout the study. Flowers contained the highest cyantraniliprole residues up to 21 DAT, then gradually decreased. Immature green fruits had the highest cyantraniliprole residues (5.3 ± 0.7 ng/g; 42 DAT), and decreased toward red ripening stages (1.4 ± 0.2 ng/g; 84 DAT). Metabolism of cyantraniliprole primarily occurred in the foliage, where 21 metabolites were tentatively identified. Flowers and fruits contained 14 and four of these metabolites, respectively. Major transformation pathways were characterized by ring closure, followed by N-demethylation, and glycosylation. Additionally, plant metabolism of cyantraniliprole was also associated with several minor phase-I, phase-II, and breakdown metabolites. The occurrence of these metabolites in plants varied as a function of tissue types and their developmental stages. Our study highlights a tissue-specific biotransformation and accumulation of metabolites of cyantraniliprole in tomato.

A starting guide to root ecology: strengthening ecological concepts and standardising root classification, sampling, processing and trait measurements.

In the context of a recent massive increase in research on plant root functions and their impact on the environment, root ecologists currently face many important challenges to keep on generating cutting-edge, meaningful and integrated knowledge. Consideration of the below-ground components in plant and ecosystem studies has been consistently called for in recent decades, but methodology is disparate and sometimes inappropriate. This handbook, based on the collective effort of a large team of experts, will improve trait comparisons across studies and integration of information across databases by providing standardised methods and controlled vocabularies. It is meant to be used not only as starting point by students and scientists who desire working on below-ground ecosystems, but also by experts for consolidating and broadening their views on multiple aspects of root ecology. Beyond the classical compilation of measurement protocols, we have synthesised recommendations from the literature to provide key background knowledge useful for: (1) defining below-ground plant entities and giving keys for their meaningful dissection, classification and naming beyond the classical fine-root vs coarse-root approach; (2) considering the specificity of root research to produce sound laboratory and field data; (3) describing typical, but overlooked steps for studying roots (e.g. root handling, cleaning and storage); and (4) gathering metadata necessary for the interpretation of results and their reuse. Most importantly, all root traits have been introduced with some degree of ecological context that will be a foundation for understanding their ecological meaning, their typical use and uncertainties, and some methodological and conceptual perspectives for future research. Considering all of this, we urge readers not to solely extract protocol recommendations for trait measurements from this work, but to take a moment to read and reflect on the extensive information contained in this broader guide to root ecology, including sections I-VII and the many introductions to each section and root trait description. Finally, it is critical to understand that a major aim of this guide is to help break down barriers between the many subdisciplines of root ecology and ecophysiology, broaden researchers' views on the multiple aspects of root study and create favourable conditions for the inception of comprehensive experiments on the role of roots in plant and ecosystem functioning.

Coordination between compound-specific chemistry and morphology in plant roots aligns with ancestral mycorrhizal association in woody angiosperms.

Recent studies on fine root functional traits proposed a root economics hypothesis where adaptations associated with mycorrhizal dependency strongly influence the organization of root traits, forming a dominant axis of trait covariation unique to roots. This conclusion, however, is based on tradeoffs of a few widely studied root traits. It is unknown how other functional traits fit into this mycorrhizal-collaboration gradient. Here, we provide a significant extension to the field of root ecology by examining how fine root secondary compounds coordinate with other root traits. We analyzed a dataset integrating compound-specific chemistry, morphology and anatomy of fine roots and leaves from 34 temperate tree species spanning major angiosperm lineages. Our data uncovered previously undocumented coordination where root chemistry, morphology and anatomy covary with each other. This coordination, aligned with mycorrhizal colonization, reflects tradeoffs between chemical protection and mycorrhizal dependency, and provides mechanistic support for the mycorrhizal-collaboration gradient. We also found remarkable phylogenetic structuring in root chemistry. These patterns were not mirrored by leaves. Furthermore, chemical protection was largely decoupled from the leaf economics spectrum. Our results unveil broad organization of root chemistry, demonstrate unique belowground adaptions, and suggest that root strategies and phylogeny could impact biogeochemical cycles through their links with root chemistry.

Nitrogen Fertilization Influences the Quantity, Composition, and Tissue Association of Foliar Phenolics in Strawberries.

Unlike quantitative changes, the compositional changes of plant phenolics and changes in their tissue association as influenced by the nutrient supply are less well understood. We evaluated the quantity, composition, and tissue association of phenolics in leaves of two Fragaria ananassa cultivars in response to different levels of nitrogen (N) fertilization using global metabolomic approaches. Influence of N supply on phenolic content in both cultivars was similar, but the magnitude of this response was compound specific. Ellagitannins, the most abundant class of phenolic oligomers, were less responsive to the applied N treatments, whereas proanthocyanidins, the less abundant class of phenolic oligomers, exhibited higher fold change. Within mono-phenolics, the hydroxycinnamates were more abundant but showed lower fold change than the hydroxybenzoates. Among flavonoids, the hydroxylated flavonols showed higher abundances than the flavones, with a preferential accumulation of dihydroxylated flavonol at lower N levels. Furthermore, glycosylated flavonols were higher than the acylated forms. The extractable fraction of phenolics was more influenced by the N treatment than the fiber-bound fraction. The extensive compositional modification of phenolics and a greater response of non-bound fractions in response to N rates highlight the potential to use precise management of N supply as an effective strategy to enhance the bioactive compounds in crops.

Genetic Architecture of Maize Rind Strength Revealed by the Analysis of Divergently Selected Populations.

The strength of the stalk rind, measured as rind penetrometer resistance (RPR), is an important contributor to stalk lodging resistance. To enhance the genetic architecture of RPR, we combined selection mapping on populations developed by 15 cycles of divergent selection for high and low RPR with time-course transcriptomic and metabolic analyses of the stalks. Divergent selection significantly altered allele frequencies of 3,656 and 3,412 single- nucleotide polymorphisms (SNPs) in the high and low RPR populations, respectively. Surprisingly, only 110 (1.56%) SNPs under selection were common in both populations, while the majority (98.4%) were unique to each population. This result indicated that high and low RPR phenotypes are produced by biologically distinct mechanisms. Remarkably, regions harboring lignin and polysaccharide genes were preferentially selected in high and low RPR populations, respectively. The preferential selection was manifested as higher lignification and increased saccharification of the high and low RPR stalks, respectively. The evolution of distinct gene classes according to the direction of selection was unexpected in the context of parallel evolution and demonstrated that selection for a trait, albeit in different directions, does not necessarily act on the same genes. Tricin, a grass-specific monolignol that initiates the incorporation of lignin in the cell walls, emerged as a key determinant of RPR. Integration of selection mapping and transcriptomic analyses with published genetic studies of RPR identified several candidate genes including ZmMYB31, ZmNAC25, ZmMADS1, ZmEXPA2, ZmIAA41 and hk5. These findings provide a foundation for an enhanced understanding of RPR and the improvement of stalk lodging resistance.

Dissipation and transformation of the diamide insecticide cyantraniliprole in ornamental snapdragon (Antirrhinum majus).

Dissipation and transformation of cyantraniliprole, a new diamide class of insecticides, were investigated under greenhouse conditions, using snapdragon (Antirrhinum majus) as the model plant. Dissipation of cyantraniliprole in treated leaves was found to be dependent upon application methods (foliar spray versus soil drench) and doses (high versus low dose), with the parent insecticide being the major residue at various sampling points. A high-dose foliar application resulted in pesticide residue of 6.7-23.8 µg/g foliar fresh weight over 8 weeks of treatments, while in soil drench treatment the residue varied from 0.8 to 1.4 µg/g. However, the residue contents were similar between the two application methods at a low application dose. The transformation pathways of cyantraniliprole were primarily intramolecular rearrangements, with IN-J9Z38 being the major metabolite across treatments. Several other metabolites were also identified, some of which were unique to the application methods. Out of total 26 metabolites tentatively identified in this study, 10 metabolites were unique to foliar application, while six metabolites were unique to soil drench. In addition to plant-mediated biotransformation, photodegradation of the parent compound was identified as a potential mechanism in foliar application.

Chemical plasticity in the fine root construct of Quercus spp. varies with root order and drought.

Fine roots of trees exhibit varying degree of plasticity to adapt to environmental stress. Although the morphological and physiological plasticity of roots has been well studied, less known are the accompanying changes in the chemical composite (chemical plasticity) of fine roots, which regulates both root function and soil carbon sequestration. We investigated the changes in quantity, composition and localization of phenolic compounds in fine root orders of Quercus alba and Quercus rubra subjected to drought stress. In both species the total quantity of lignins varied only by root orders, where the distal (first and second) root orders had lower lignin compared to higher orders. Despite a lower lignin content, the distal root orders had higher content of guaiacyl lignin and bound phenolics that would provide a greater meshing of lignocellulosic matrix, and thus a higher tissue integrity. Unlike lignins, drought altered the quantity and composition of tannins. In Q. alba, the ellagitannins decreased in the distal root orders exposed to drought, while the fiber-bound condensed tannnins increased. The lower content of ellagitannins with antimicrobial properties under drought reveals an adaptive response by fine roots to promote symbiotic association, as evidenced by the higher colonization of ectomycorrhizal fungi. Our study revealed that, when exposed to drought, the composition of heteropolymers are strategically varied across fine root orders, so as to provide a greater root function without compromising the tissue protection.

Proteasome Inhibition in Brassica napus Roots Increases Amino Acid Synthesis to Offset Reduced Proteolysis.

Cellular homeostasis is maintained by the proteasomal degradation of regulatory and misfolded proteins, which sustains the amino acid pool. Although proteasomes alleviate stress by removing damaged proteins, mounting evidence indicates that severe stress caused by salt, metal(oids), and some pathogens can impair the proteasome. However, the consequences of proteasome inhibition in plants are not well understood and even less is known about how its malfunctioning alters metabolic activities. Lethality causes by proteasome inhibition in non-photosynthetic organisms stem from amino acid depletion, and we hypothesized that plants respond to proteasome inhibition by increasing amino acid biosynthesis. To address these questions, the short-term effects of proteasome inhibition were monitored for 3, 8 and 48 h in the roots of Brassica napus treated with the proteasome inhibitor MG132. Proteasome inhibition did not affect the pool of free amino acids after 48 h, which was attributed to elevated de novo amino acid synthesis; these observations coincided with increased levels of sulfite reductase and nitrate reductase activities at earlier time points. However, elevated amino acid synthesis failed to fully restore protein synthesis. In addition, transcriptome analysis points to perturbed abscisic acid signaling and decreased sugar metabolism after 8 h of proteasome inhibition. Proteasome inhibition increased the levels of alternative oxidase but decreased aconitase activity, most sugars and tricarboxylic acid metabolites in root tissue after 48 h. These metabolic responses occurred before we observed an accumulation of reactive oxygen species. We discuss how the metabolic response to proteasome inhibition and abiotic stress partially overlap in plants.