Salicylic Acid-Induced Elicitation of Nepetalactone and Rosmarinic Acid Biosynthesis in Naked Catmint (Nepeta nuda L.): Metabolomic and Transcriptional Insights.

Salicylic acid (SA) is involved in plant defense responses to environmental stressors by modulating gene expression and specialized metabolites production, enhancing plant adaptive resilience through systemic signaling pathways. This study investigates the impact of exogenous application of SA on the metabolism of iridoids and phenolic compounds-characteristic specialized metabolites of the Nepeta species, associated with diverse biological activities. Nepetalactone (NL) is a characteristic monoterpene iridoid, while rosmarinic acid (RA) represents the most abundant phenolic compound within the genus. We explored the effects of varying SA concentrations (2 µM, 5 µM, 10 µM, and 20 µM) on iridoid and phenolic metabolism in in vitro-grown Nepeta nuda, following 7 days and 28 days of elicitation. A significant increase in trans,trans-NL content was observed after 7-day exposure to 2 µM SA, while prolonged exposure led to a decrease in its levels, particularly at higher SA concentrations. Gene expression analysis revealed that 7 days of exposure to lower concentrations of SA upregulated genes coding for NAD-dependent nepetalactol-related short-chain dehydrogenase/reductases (NEPSs), key regulatory enzymes catalyzing the final steps of NL biosynthesis. In contrast, prolonged exposure to 20 µM SA downregulated genes coding for geraniol 8-hydroxylase (NnG8H) and 8-hydroxygeraniol oxidoreductase (Nn8HGO), which resulted in reduced iridoid content. Conversely, SA treatment notably increased RA content after prolonged exposure to 20 µM SA, which is a result of the enhanced expression of all analyzed RA biosynthesis-related genes. These findings demonstrate that both concentration and duration of SA treatment are critical determinants of elicitation outcomes in N. nuda. Strategic manipulation of these parameters can redirect metabolic flux toward either iridoid or phenolic compounds production, and enhance biotechnological production of specialized metabolites in N. nuda.