Methyl Jasmonate Activates the 2C Methyl-D-erithrytol 2,4-cyclodiphosphate Synthase Gene and Stimulates Tanshinone Accumulation in Salvia miltiorrhiza Solid Callus Cultures

• Published in: Molecules (Basel, Switzerland) Switzerland), 2022-03, Vol.27 (6), p.1772
• Main Authors: Szymczyk, Piotr, Szymańska, Grażyna, Kuźma, Łukasz, Jeleń, Agnieszka, Balcerczak, Ewa
• Format: Article
• Language: English
• Subjects: Abietanes; Accumulation; Acetates; Alzheimer's disease; Biosynthesis; Callus; callus culture; Cardiovascular disease; cis-active element; Cryptotanshinone; Cyclopentanes - metabolism; Cyclopentanes - pharmacology; Enzymes; Gene expression; Gene regulation; MEP pathway; Methyl jasmonate; Nucleotides; Oxylipins - metabolism; Oxylipins - pharmacology; Polymerase chain reaction; promoter; Salvia miltiorrhiza; Signal transduction; tanshinone; Tanshinones; Transcription factors; Transcriptomics
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules27061772

The present study characterizes the 5' regulatory region of the gene. The isolated fragment is 1559 bp long and consists of a promoter, 5'UTR and 31 nucleotide 5' fragments of the CDS region. In silico bioinformatic analysis found that the promoter region contains repetitions of many potential -active elements. -active elements associated with the response to methyl jasmonate (MeJa) were identified in the gene promoter. Co-expression studies combined with earlier transcriptomic research suggest the significant role of MeJa in gene regulation. These findings were in line with the results of the RT-PCR test showing gene expression induction after 72 h of MeJa treatment. Biphasic total tanshinone accumulation was observed following treatment of solid callus cultures with 50-500 μM methyl jasmonate, with peaks observed after 10-20 and 50-60 days. An early peak of total tanshinone concentration (0.08%) occurred after 20 days of 100 μM MeJa induction, and a second, much lower one, was observed after 50 days of 50 μM MeJa stimulation (0.04%). The dominant tanshinones were cryptotanshinone (CT) and dihydrotanshinone (DHT). To better understand the inducing effect of MeJa treatment on tanshinone biosynthesis, a search was performed for methyl jasmonate-responsive -active motifs in the available sequences of gene proximal promoters associated with terpenoid precursor biosynthesis. The results indicate that MeJa has the potential to induce a significant proportion of the presented genes, which is in line with available transcriptomic and RT-PCR data.