Nitrate modulates pectin metabolism and cell wall mechanics during cell expansion in Arabidopsis

Nitrate is a key nutrient and one of the most important nitrogen sources for land plants. Besides its nutritional role, nitrate is a signal molecule that regulates plant gene expression, metabolism, physiology, growth, and development. In cotyledons and true leaves, nitrate promotes growth by inducing cell expansion. Plant cell expansion requires changes in the cell wall. However, there is scant information on the influence of nitrate on cell wall metabolism and properties during cell expansion and growth. Here, we demonstrate that nitrate availability modulates pectin metabolism, a major polysaccharide of the primary cell wall. Using colorimetric assays, immunohistochemistry, and confocal microscopy, we show that nitrate enhances methylesterified pectin during cotyledon cell expansion. This is achieved by increasing galacturonic acid (GalA) deposition as homogalacturonan (HG) and by decreasing global PME activity. We further show that this regulation is dependent on nitrate signaling pathway components, including NRT1.1 and NLP7. Pectin methylesterification state impacts the mechanical properties of the cell wall. We characterized cell wall elasticity changes during nitrate-induced expansion using atomic force microscopy (AFM) and automatic confocal microextensometry (ACME). We found that nitrate induces cell wall softening at both cellular and whole-tissue levels during this expansion process. Our results indicate pectin metabolism plays an important role in nitrate-induced cell expansion and cotyledon growth in Arabidopsis. We provide insights into the interplay between nitrate signaling, cell wall metabolism, and biomechanical properties for cell expansion. Our results contribute to our understanding of how plants sense and respond to environmental cues for growth.