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We showed previously that nitrogen (N) limitation decreases Arabidopsis resistance to Erwinia amylovora (Ea). We show that decreased resistance to bacteria in low N is correlated with lower apoplastic reactive oxygen species (ROS) accumulation and lower jasmonic acid (JA) pathway expression. Consistently, pretreatment with methyl jasmonate (Me‐JA) increased the resistance of plants grown under low N. In parallel, we show that in planta titres of a nonvirulent type III secretion system (T3SS)‐deficient Ea mutant were lower than those of wildtype Ea in low N, as expected, but surprisingly not in high N. This lack of difference in high N was consistent with the low expression of the T3SS‐encoding hrp virulence genes by wildtype Ea in plants grown in high N compared to plants grown in low N. This suggests that expressing its virulence factors in planta could be a major limiting factor for Ea in the nonhost Arabidopsis. To test this hypothesis, we preincubated Ea in an inducing medium that triggers expression of hrp genes in vitro, prior to inoculation. This preincubation strongly enhanced Ea titres in planta, independently of the plant N status, and was correlated to a significant repression of JA‐dependent genes. Finally, we identify two clusters of metabolites associated with resistance or with susceptibility to Ea. Altogether, our data showed that high susceptibility of Arabidopsis to Ea, under low N or following preincubation in hrp‐inducing medium, is correlated with high expression of the Ea hrp genes in planta and low expression of the JA signalling pathway, and is correlated with the accumulation of specific metabolites. [ABSTRACT FROM AUTHOR]

SUMMARY: The SID2 (SA INDUCTION‐DEFICIENT2) gene that encodes ICS1 (isochorismate synthase), plays a central role in salicylic acid biosynthesis in Arabidopsis. The sid2 and NahG (encoding a bacterial SA hydroxylase) overexpressing mutants (NahG‐OE) have currently been shown to outperform wild type, presenting delayed leaf senescence, higher plant biomass and better seed yield. When grown under sulfate‐limited conditions (low‐S), sid2 mutants exhibited early leaf yellowing compared to the NahG‐OE, the npr1 mutant affected in SA signaling pathway, and WT. This indicated that the hypersensitivity of sid2 to sulfate limitation was independent of the canonical npr1 SA‐signaling pathway. Transcriptomic and proteomic analyses revealed that major changes occurred in sid2 when cultivated under low‐S, changes that were in good accordance with early senescence phenotype and showed the exacerbation of stress responses. The sid2 mutants displayed a lower sulfate uptake capacity when cultivated under low‐S and lower S concentrations in their rosettes. Higher glutathione concentrations in sid2 rosettes under low‐S were in good accordance with the higher abundance of proteins involved in glutathione and ascorbate redox metabolism. Amino acid and lipid metabolisms were also strongly modified in sid2 under low‐S. Depletion of total fatty acids in sid2 under low‐S was consistent with the fact that S‐metabolism plays a central role in lipid synthesis. Altogether, our results show that functional ICS1 is important for plants to cope with S limiting conditions. [ABSTRACT FROM AUTHOR]

Esca is a complex grapevine trunk disease caused by wood‐rotting ascomycetes and basidiomycetes and leading to several foliar and wood symptoms. Given that the esca expression can be influenced by several environmental, physiological, and genetic factors, foliar symptoms are inconsistent in incidence and prevalence and may appear 1 year but not the following. We have previously reported a clone‐dependent expression of the disease in cv Chardonnay. Owing to metabolome analysis, we could discriminate the metabolite fingerprint of green leaves collected on diseased vines of clones 76 and 95. These clone‐dependent fingerprints were year‐dependent in intensity and nature. The present work was conducted to determine if the clone‐dependent disease expression observed is specific to Chardonnay or if it also occurs in another cultivar. A plot located in the Jura vineyard (France) and planted with both 1004 and 1026 clones of Trousseau, a cultivar highly susceptible to esca, was thus selected and studied during 2017 and 2018. A year‐dependent variation of the symptoms expression was first observed and a possible relationship with rainfall is hypothesized and discussed. Moreover, a higher percentage of the clone 1026 vines expressed disease, compared to the 1004 ones, suggesting the higher susceptibility of this clone. Finally, metabolomic analyses of the remaining green leaves (i.e, without symptom expression) of partial esca‐apoplectic vines allowed us to confirm a clone‐dependent metabolic response to the disease. The metabolite fingerprints obtained differed in nature and intensity to those previously reported for Chardonnay and also between years. [ABSTRACT FROM AUTHOR]

During leaf senescence, nitrogen is remobilized and carbon backbones are replenished by amino acid catabolism, with many of the key reactions occurring in mitochondria. The intermediate Δ1‐pyrroline‐5‐carboxylate (P5C) is common to some catabolic pathways, thus linking the metabolism of several amino acids, including proline and arginine. Specifically, mitochondrial proline catabolism involves sequential action of proline dehydrogenase (ProDH) and P5C dehydrogenase (P5CDH) to produce P5C and then glutamate. Arginine catabolism produces urea and ornithine, the latter in the presence of α‐ketoglutarate being converted by ornithine δ‐aminotransferase (OAT) into P5C and glutamate. Metabolic changes during dark‐induced leaf senescence (DIS) were studied in Arabidopsis thaliana leaves of Col‐0 and in prodh1prodh2, p5cdh and oat mutants. Progression of DIS was followed by measuring chlorophyll and proline contents for 5 days. Metabolomic profiling of 116 compounds revealed similar profiles of Col‐0 and oat metabolism, distinct from prodh1prodh2 and p5cdh metabolism. Metabolic dynamics were accelerated in p5cdh by 1 day. Notably, more P5C and proline accumulated in p5cdh than in prodh1prodh2. ProDH1 enzymatic activity and protein amount were significantly down‐regulated in p5cdh mutant at Day 4 of DIS. Mitochondrial P5C levels appeared critical in determining the flow through interconnected amino acid remobilization pathways to sustain senescence. Summary statement: Metabolic profiling revealed a key role of proline dehydrogenase (ProDH) and pyrroline‐5‐carboxylate dehydrogenase (P5CDH) but not of ornithine aminotransferase in proline oxidation during Arabidopsis dark‐induced leaf senescence (DIS). P5CDH‐dependent regulation of ProDH appears to be necessary to partition the limited resources during DIS to ensure the Krebs cycle to be fuelled with TCA intermediates for remobilization of the remaining nitrogen in the leaf. [ABSTRACT FROM AUTHOR]

Despite a general view that asparagine synthetase generates asparagine as an amino acid for long-distance transport of nitrogen to sink organs, its role in nitrogen metabolic pathways in floral organs during seed nitrogen filling has remained undefined. We demonstrate that the onset of pollination in Arabidopsis induces selected genes for asparagine metabolism, namely ASN1 ( At3g47340), GLN2 ( At5g35630), GLU1 ( At5g04140), Aap AT2 ( At5g19950), ASPGA1 ( At5g08100) and ASPGB1 ( At3g16150), particularly at the ovule stage (stage 0), accompanied by enhanced asparagine synthetase protein, asparagine and total amino acids. Immunolocalization confined asparagine synthetase to the vascular cells of the silique cell wall and septum, but also to the outer and inner seed integuments, demonstrating the post-phloem transport of asparagine in these cells to developing embryos. In the asn1 mutant, aberrant embryo cell divisions in upper suspensor cell layers from globular to heart stages assign a role for nitrogen in differentiating embryos within the ovary. Induction of asparagine metabolic genes by light/dark and nitrate supports fine shifts of nitrogen metabolic pathways. In transgenic Arabidopsis expressing promoter Ca MV 35S:: ASN1 fusion, marked metabolomics changes at stage 0, including a several-fold increase in free asparagine, are correlated to enhanced seed nitrogen. However, specific promoter Napin2S:: ASN1 expression during seed formation and a six-fold increase in asparagine toward the desiccation stage result in wild-type seed nitrogen, underlining that delayed accumulation of asparagine impairs the timing of its use by releasing amide and amino nitrogen. Transcript and metabolite profiles in floral organs match the carbon and nitrogen partitioning to generate energy via the tricarboxylic acid cycle, GABA shunt and phosphorylated serine synthetic pathway. [ABSTRACT FROM AUTHOR]

ABSTRACT We investigated the function of ASN2, one of the three genes encoding asparagine synthetase (EC 6.3.5.4), which is the most highly expressed in vegetative leaves of Arabidopsis thaliana. Expression of ASN2 and parallel higher asparagine content in darkness suggest that leaf metabolism involves ASN2 for asparagine synthesis. In asn2-1 knockout and asn2-2 knockdown lines, ASN2 disruption caused a defective growth phenotype and ammonium accumulation. The asn2 mutant leaves displayed a depleted asparagine and an accumulation of alanine, GABA, pyruvate and fumarate, indicating an alanine formation from pyruvate through the GABA shunt to consume excess ammonium in the absence of asparagine synthesis. By contrast, asparagine did not contribute to photorespiratory nitrogen recycle as photosynthetic net CO2 assimilation was not significantly different between lines under both 21 and 2% O2. ASN2 was found in phloem companion cells by in situ hybridization and immunolocalization. Moreover, lack of asparagine in asn2 phloem sap and lowered 15N flux to sinks, accompanied by the delayed yellowing (senescence) of asn2 leaves, in the absence of asparagine support a specific role of asparagine in phloem loading and nitrogen reallocation. We conclude that ASN2 is essential for nitrogen assimilation, distribution and remobilization ( via the phloem) within the plant. [ABSTRACT FROM AUTHOR]

Inactivation of Arabidopsis WAT1 (Walls Are Thin1), a gene required for secondary cell-wall deposition, conferred broad-spectrum resistance to vascular pathogens, including the bacteria Ralstonia solanacearum and Xanthomonas campestris pv. campestris, and the fungi Verticillium dahliae and Verticillium albo-atrum. Introduction of NahG, the bacterial salicylic acid (SA)-degrading salicylate hydroxylase gene, into the wat1 mutant restored full susceptibility to both R. solanacearum and X. campestris pv. campestris. Moreover, SA content was constitutively higher in wat1 roots, further supporting a role for SA in wat1-mediated resistance to vascular pathogens. By combining transcriptomic and metabolomic data, we demonstrated a general repression of indole metabolism in wat1-1 roots as shown by constitutive down-regulation of several genes encoding proteins of the indole glucosinolate biosynthetic pathway and reduced amounts of tryptophan (Trp), indole-3-acetic acid and neoglucobrassicin, the major form of indole glucosinolate in roots. Furthermore, the susceptibility of the wat1 mutant to R. solanacearum was partially restored when crossed with either the trp5 mutant, an over-accumulator of Trp, or Pro35S:AFB1-myc, in which indole-3-acetic acid signaling is constitutively activated. Our original hypothesis placed cell-wall modifications at the heart of the wat1 resistance phenotype. However, the results presented here suggest a mechanism involving rootlocalized metabolic channeling away from indole metabolites to SA as a central feature of wat1 resistance to R. solanacearum. [ABSTRACT FROM AUTHOR]