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Siffert, A.

Publications and source records attributed to Siffert, A..

3 recordsLinked to original sources

Altered Arabidopsis thaliana sugar metabolism affects exudation, immune responses, and plant-microbe interactions

Sugars are critical for plant growth, development, and environmental interactions. They have multiple roles as nutrients for plants, associated beneficial and pathogenic microbes, and as signaling compounds for immunity. We characterize the interconnectedness of these functions by analyzing sugar metabolism and transporter mutant lines. We find that in these lines, root-derived compounds, exudates, are significantly altered in comparison with wild-type not only for carbohydrates, but also for lipids, organic acids, and defense compounds. Quantification of sugar exudation reveals more carbon release during the day than at night, altered sugar exudation in mutant lines, and an opposite exudation pattern with elevated exudation at night for pgm1, a line deficient in starch synthesis. Sugar levels in exudates and tissues did not correlate, suggesting a controlled mode of exudation for sugars. Altered sugar levels have functional consequences: mutant lines exhibit increased resistance against the pathogen Pseudomonas syringae and harbor altered numbers of microbes on roots. Day- and nighttime exudates of mutant lines impact the growth of single microbes such as an inability to grow for Bacillus subtilis. Exogenous sugar alters the production of reactive oxygen species in a plant development-dependent manner with opposite effects at 9 days and 14 days. An RNAseq experiment reveals candidate genes potentially involved in this regulation. Our data highlight that sugar metabolism is intricately linked with other metabolite pathways. Alteration of single genes in central carbon metabolism profoundly alters plant immune responses and plant-microbe interactions.

plant biology↗

Distinct metabolite profiles in tissues and exudates of a monocot and dicot shaped by the environment

Plants exhibit remarkable plasticity in response to environmental changes. Understanding how plants adapt to diverse environmental conditions through changes in their metabolite profiles can provide insights into their adaptive strategies under suboptimal climate conditions. For this, metabolite profiles of tissues and root-derived, exuded compounds in various environmental conditions need to be characterized. Here, we compare the shoot, root, and root exudate metabolite profiles of the monocot Brachypodium distachyon and the dicot Arabidopsis thaliana grown in sterile, non-sterile, and sucrose-supplied basal salt medium or soil extract to represent natural and various standard laboratory conditions. We report unique metabolite fingerprints in shoots and roots for each species and environmental condition. Exuded compounds of Arabidopsis displayed higher sensitivity to soil extract conditions, whereas Brachypodium showed significant changes in response to non-sterile conditions. Organic acids, lipids, organic oxygen compounds, and phenylpropanoids were major contributors to the observed differences. Our results highlight the importance of considering environmental aspects when investigating plant metabolism and point towards crucial chemical classes involved in plant-microbe-environment interactions.

plant biology↗

A key residue of the extracellular gate provides quality control contributing to ABCG substrate specificity

For G-type ATP-binding cassette (ABC) transporters, a hydrophobic "di-leucine motif" as part of a hydrophobic extracellular gate has been described to separate a large substrate-binding cavity from a smaller upper cavity and proposed to act as a valve controlling drug extrusion. Here, we show that an L704F mutation in the hydrophobic extracellular gate of Arabidopsis ABCG36/PDR8/PEN3 uncouples the export of the auxin precursor indole-3-butyric acid (IBA) from that of the defense compound camalexin (CLX). Molecular dynamics simulations reveal an increase in free energy and pulling forces for CLX at both the entrance and exit sites of ABCG36L704F, respectively, providing a mechanistic rationale for the transport discrimination of CLX. Mutagenesis of L704 to tyrosine allows export of structurally related non-ABCG36 substrates, indole-3-acetic acid (IAA) and indole, suggesting an allosteric communication between the extracellular gate and the central substrate binding pocket. In summary, our work supports the conclusion that L704 is a key residue of the extracellular gate that provides a final quality control contributing to ABCG substrate specificity.

biochemistry↗