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Boerjan, W.

Publications and source records attributed to Boerjan, W..

7 recordsLinked to original sources

The Arabidopsis PAL3 is a carboxy-tyrosine ammonia lyase

Phenylalanine ammonia lyase (PAL) catalyses the deamination of L-phenylalanine, which is the first step of the plant-specific phenylpropanoid pathway. Its position at the intersection of primary and specialized metabolism, combined with its important role in plant growth and adaptive stress response, has made it a subject of extensive studies. We identified key amino acids in the catalytic pocket of several PAL enzymes, including PAL3 of Arabidopsis, that are different from the canonical PAL sites, suggesting these enzymes have a different substrate specificity and have been miscategorised for decades. By combining untargeted metabolomics with enzyme assays, we discovered that PAL3 converts 3-carboxy-tyrosine into carboxy-p-coumaric acid. Based on this specific function, we propose to denote it as a 3-carboxy-tyrosine ammonia lyase (CAL). In addition to its discovery as a novel enzyme class, paving the way for biotechnological applications, we introduce the carboxy-phenylpropanoids as a new class of specialised metabolites.

biochemistry↗

Metabolic engineering of a tyrosine-specific phenylpropanoid pathway in plants

While all plants use L-phenylalanine for phenylpropanoid biosynthesis, grasses can also initiate the pathway from L-tyrosine. Curiously, no plant has evolved an exclusive tyrosine-derived route. We generate plants with phenylpropanoid biosynthesis initiated from phenylalanine, tyrosine, or both by expressing a Brachypodium phenylalanine/tyrosine ammonia-lyase (PTAL) in Arabidopsis WT and c4h mutants. Engineering a bifunctional phenylpropanoid pathway in WT plants did not negatively impact growth, while introducing a tyrosine-specific pathway in the c4h mutant could overcome the seedling-lethal phenotype. Interestingly, restored c4h mutants relying solely on the tyrosine route displayed developmental defects linked to the strong overaccumulation of the auxin transport inhibitor cis-cinnamic acid. Our findings suggest that the requirement of this widely overlooked plant metabolite could be the crucial factor for the evolutionary retention of the canonical phenylpropanoid biosynthesis route via L-phenylalanine in plants.

plant biology↗

Introducing furanocoumarin biosynthetic genes in tomato results in coumarins accumulation and impacted growth

Over the past three decades, eeorts to decipher plant metabolism have shed light on key enzymes driving specialized metabolite biosynthesis. Although only few pathways have been completely investigated to date, their characterization paves the way for exploring the potential eeects of specialized metabolites on plant physiology. Among them is the linear furanocoumarin pathway, which was recently completed to produce up to psoralen. In this study, we report the first metabolic engineering of the linear furanocoumarin pathway to enable artificial psoralen production in tomato, through the integration of four genes coding for the enzymes: Umbelliferone Synthase, Demethylsuberosin Synthase, Marmesin Synthase and Psoralen Synthase. Interestingly, coumarins were produced instead of furanocoumarins. Using morphophysiological, metabolomic, and transcriptomic analyses, we suggest how coumarins, particularly scopoletin, can impact growth and aeect plant physiology, even at low concentrations. As coumarins have increasingly attracted interest for agricultural applications due to their minimal environmental impact, this work both expands and challenges their potential by highlighting the physiological costs and benefits they may impose on tomato. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=164 SRC="FIGDIR/small/663522v2_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@1df0bdeorg.highwire.dtl.DTLVardef@843d9borg.highwire.dtl.DTLVardef@1e898b6org.highwire.dtl.DTLVardef@18205eb_HPS_FORMAT_FIGEXP M_FIG Graphical abstract C_FIG

plant biology↗

Genetic architecture of the tomato fruit lipidome; new insights link lipid and volatile compounds

Tomato (Solanum lycopersicum L.) fruit flavor is determined by a combination of multiple volatile compounds, including several derived from lipids and fatty acids. Although fruit flavor has been intensively studied, the linkage between lipid metabolism and flavor remains largely undefined. Here, we performed a genome-wide association study (GWAS) and QTL mapping for the fruit lipid content from 550 tomato accessions and 107 backcross inbred lines (BILs) in two consecutive seasons. Over 130 lipid compounds were identified and mapped, allowing for the identification of over 600 metabolic QTL (mQTL). We further described and validated candidate genes associated with lipid content. Among them is a lipase-like protein (TomLLP) whose function was validated in vivo using overexpression lines in tomato and knockout mutants in Arabidopsis. We also identified functions for three enzymes: a class III lipase (Sl-LIP8), a cyclopropane-fatty-acyl-phospholipid synthase (CFAPS1), and Lipoxygenase C (TomLoxC). By utilizing knockout lines for CFAPS1 and CRISPR-Cas9 loss-of-function lines for Sl-LIP8 and TomLoxC, we demonstrated the functional importance of these enzymes in fruit lipid metabolism. Our study provides a comprehensive analysis of the tomato fruit lipidome and insights into key genes that shaped the natural variation in tomato lipid content and their links to flavor-associated volatile compounds.

molecular biology↗

NON-SPECIFIC EFFECTS OF A CINNAMATE-4-HYDROXYLASE INHIBITOR ON AUXIN HOMEOSTASIS

Chemical inhibitors are often implemented for the functional characterization of genes to overcome the limitations associated with genetic approaches. Although being a powerful tool, off-target effects of these inhibitors are easily overlooked in a complex biological setting. Here we illustrate the implications of such secondary effects by focusing on piperonylic acid (PA), an inhibitor of CINNAMATE-4-HYDROXYLASE (C4H) that is often used to investigate the involvement of lignin during plant growth and development. When supplied to plants, we found that PA is recognized as a substrate by GRETCHEN HAGEN 3.6 (GH3.6), an amido synthetase involved in the formation of the auxin catabolite indole-3-acetic acid (IAA)-Asp. By competing for the same enzyme, PA interferes with auxin conjugation, resulting in an increase in cellular auxin concentrations. These increased auxin levels likely further contribute to an increase in adventitious rooting previously observed upon PA-treatment. Despite the focus on GH3.6 in this report, PA is conjugated by an array of enzymes and their subsequent reduced activity on native substrates could potentially affect a whole set of physiological processes in the plant. We conclude that surrogate occupation of the endogenous conjugation machinery in the plant by exogenous compounds is likely a more general phenomenon that is rarely considered in pharmacological studies. Our results hereby provide an important basis for future reference in studies using chemical inhibitors.

plant biology↗

Schengen-pathway controls spatially separated and chemically distinct lignin deposition in the endodermis.

Lignin is a complex polymer precisely deposited in the cell wall of specialised plant cells, where it provides essential cellular functions. Plants coordinate timing, location, abundance and composition of lignin deposition in response to endogenous and exogenous cues. In roots, a fine band of lignin, the Casparian strip encircles endodermal cells. This forms an extracellular barrier to solutes and water and plays a critical role in maintaining nutrient homeostasis. A signalling pathway senses the integrity of this diffusion barrier and can induce over-lignification to compensate for barrier defects. Here, we report that activation of this endodermal sensing mechanism triggers a transcriptional reprogramming strongly inducing the phenylpropanoid pathway and immune signaling. This leads to deposition of compensatory lignin that is chemically distinct from Casparian strip lignin. We also report that a complete loss of endodermal lignification drastically impacts mineral nutrients homeostasis and plant growth.

plant biology↗

The genome and metabolome of the tobacco tree, Nicotiana glauca: a potential renewable feedstock for the bioeconomy

BackgroundGiven its tolerance to stress and its richness in particular secondary metabolites, the tobacco tree, Nicotiana glauca, has been considered a promising biorefinery feedstock that would not be competitive with food and fodder crops.\n\nResultsHere we present a 3.5 Gbp draft sequence and annotation of the genome of N. glauca spanning 731,465 scaffold sequences, with an N50 size of approximately 92 kbases. Furthermore, we supply a comprehensive transcriptome and metabolome analysis of leaf development comprising multiple techniques and platforms.\n\nThe genome sequence is predicted to cover nearly 80% of the estimated total genome size of N. glauca. With 73,799 genes predicted and a BUSCO score of 94.9%, we have assembled the majority of gene-rich regions successfully. RNA-Seq data revealed stage-and/or tissue-specific expression of genes, and we determined a general trend of a decrease of tricarboxylic acid cycle metabolites and an increase of terpenoids as well as some of their corresponding transcripts during leaf development.\n\nConclusionThe N. glauca draft genome and its detailed transcriptome, together with paired metabolite data, constitute a resource for future studies of valuable compound analysis in tobacco species and present the first steps towards a further resolution of phylogenetic, whole genome studies in tobacco.

plant biology↗