Developmental regulation and physical interaction among enzymes involved in sorgoleone biosynthesis.

Ammonium in the soil is converted into nitrate by nitrifying bacteria or archaea. While nitrate is readily available for plants, it is prone to leaching and contributes to eutrophication. In addition, when the soil conditions become anaerobic, nitrate can be reduced to nitrous oxide, a powerful greenhouse gas. Therefore, slowing nitrification in agricultural soil offers some benefits by reducing nitrogen loss and decreasing water and air pollution. Since nitrogen is a limiting nutrient for most ecological niches, many plants have evolved specialized compounds that reduce nitrification. One such compound, sorgoleone, which is secreted from the root hair of sorghum, has been relatively well studied due to its allelopathic function, with most enzymes involved in its biosynthesis elucidated. However, the secretion mechanisms remain unknown. Previous studies reported numerous lipidic vesicles in the sorghum root hair and speculated that they are involved in sorgoleone storage or secretion, but their roles remain unclear. Also, the subcellular organelles that are involved in sorgoleone synthesis have not been identified. In the present study, we found that the expression of sorgoleone biosynthesis enzymes is induced in a specific root zone, indicating that the secretion is developmentally regulated. The accumulation of internal vesicles preceded the peak of sorgoleone biosynthesis and secretion, indicating that the vesicles play a role in precursor storage rather than secretion. Moreover, our data suggest that enzymes that catalyze the first three steps, SbDES2, SbDES3, and SbARS1, interact with each other to form a multi-enzyme complex on the endoplasmic reticulum surface.

Increased Expression of UMAMIT Amino Acid Transporters Results in Activation of Salicylic Acid Dependent Stress Response.

In addition to their role in the biosynthesis of important molecules such as proteins and specialized metabolites, amino acids are known to function as signaling molecules through various pathways to report nitrogen status and trigger appropriate metabolic and cellular responses. Moreover, changes in amino acid levels through altered amino acid transporter activities trigger plant immune responses. Specifically, loss of function of major amino acid transporter, over-expression of cationic amino acid transporter, or over-expression of the positive regulators of membrane amino acid export all lead to dwarfed phenotypes and upregulated salicylic acid (SA)-induced stress marker genes. However, whether increasing amino acid exporter protein levels lead to similar stress phenotypes has not been investigated so far. Recently, a family of transporters, namely USUALLY MULTIPLE ACIDS MOVE IN AND OUT TRANSPORTERS (UMAMITs), were identified as amino acid exporters. The goal of this study was to investigate the effects of increased amino acid export on plant development, growth, and reproduction to further examine the link between amino acid transport and stress responses. The results presented here show strong evidence that an increased expression of UMAMIT transporters induces stress phenotypes and pathogen resistance, likely due to the establishment of a constitutive stress response via a SA-dependent pathway.