Salvia abrotanoides (Kar.) Sytsma is a medicinal plant that contains phenolic acids, especially rosmarinic acid (RA), with a wide variety of health benefits. In this research, the stimulatory effects of sodium nitroprusside (SNP), a donor of nitric oxide (NO), were examined on phenolic acids accumulation in the shoot cultures of the species after different times of exposure. The developed shoots on single-nodal explants in Murashige and Skoog solid medium supplemented with 0.5 mg L−1 kinetin and 0.05 mg L−1 indole-3-acetic acid were treated with SNP at concentrations of 25, 50 and 100 μM. The highest content of RA (10.45 ± 0.40 mg g−1 DW) was measured 144 h after elicitation of the shoots with 100 μM SNP. The maximum values of salvianolic acid A (Sal-A) (0.065 ± 0.00 mg g−1 DW) and salvianolic acid B (Sal-B) (0.42 ± 0.01 mg g− 1 DW) were obtained in the shoots after 96 and 144 h exposure to 50 and 25 μM SNP, respectively. Also, elicitation with SNP at different concentrations significantly upregulated the crucial genes (PAL, TAT , RAS and CYP98A14) involved in phenolic acids biosynthesis in the shoots with distinct patterns, although no strong correlations were observed between transcription levels of the genes and phenolic acids accumulation. The findings of this study provide beneficial information about the impact of NO as an effective elicitor, which could be valuable for the in vitro improvement of phenolic acids production in S. abrotanoides.

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