Elicitation of solid callus cultures of Salvia miltiorrhiza Bunge with salicylic acid and a synthetic auxin (1-naphthaleneacetic acid)

Abstract

The presented study analyses the influence of salicylic acid (SA) and the synthetic auxin 1-naphthaleneacetic acid (NAA) on total tanshinone level and on dihydrotanshinone (DHT), cryptotanshinone (CT), tanshinine I (TI) and tanshinone IIA (TIIA) level in Salvia miltiorrhiza callus cultures growing on solid Murashige and Skoog (MS) medium. The influence of SA and NAA was evaluated at 10-day intervals throughout a 80-day treatment period. SA was applied at 0.1, 0.2 and 0.4 mM, and NAA at 2.69, 13.43, 26.85 and 40.28 μM. DHT, CT, TI and TIIA concentrations were measured using HPLC. NAA did not increase the concentration of any tanshinone. SA increased content in a concentration- and time-dependent manner; however, the yields were relatively low, possibly due to the metabolic specificity of S. miltiorrhiza cultivars in Poland. Total tanshinone concentration reached 226.38 ± 37.33 μg g−1 DW after 50 days of 0.4 mM SA elicitation. After 50 days of SA elicitation, the following maximum tanshinone concentrations were observed for 0.4 mM SA: DHT (71.58 ± 12.72 μg g−1 DW), CT (108.54 ± 18.29 μg g−1 DW), TI (29.50 ± 4.13 μg g−1 DW) and TIIA (16.75 ± 2.74 μg g−1 DW). To account for these observed differences in tanshinone biosynthesis, the distribution of SA and auxin responsive cis-active motif in the proximal promoters of the mevalonic acid, methylerythritol-4-phosphate and tanshinone-precursor biosynthesis pathway genes was evaluated in A. thaliana and S. miltiorrhiza. Our findings indicate that the SA-responsive cis-active elements have a much broader distribution than those recognized by auxin-responsive transcription factors.

Distinction of chia varieties in vivo and in vitro based on the flow cytometry and rosmarinic acid production

“…This research has confirmed that the use of elicitors in the appropriate concentration can significantly contribute to increase rosmarinic acid production. The positive effect of elicitors on the production of secondary metabolites was also confirmed by other studies conducted on in vitro cultures of various species of Salvia (Attaran Dowom et al 2022 ; Grzegorczyk-Karolak et al 2022 ; Radomir et al 2023 ; Rostami et al 2022 ; Shoja et al 2022 ; Szymczyk et al 2021 ). The stimulation of rosmarinic acid production and accumulation by elicitors such as YeE has also been observed in the cell cultures of other plant species, including Orthosiphon aristatus (Mizukami et al 1992 ), Lithospermum erythrorhizon (Sumaryono et al 1991 ; Mizukami et al 1993 ), Coleus blumei (Szabo et al 1999 ), Zingiber officinale callus cultures (Ali et al 2018 ), and Polygonum multiflorum beard root cultures (Ho et al 2018 ).…”

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

“…To give a more accurate picture of the MeJa induction of tanshinone biosynthesis in S. miltiorrhiza solid callus cultures, the distribution of the cis -active elements was analysed within the proximal promoters of MEP and MVA and the later stages of tanshinone precursor biosynthesis genes in A. thaliana and S. miltiorrhiza [ 38 ]. The obtained data were compared with available transcriptomic studies of S. miltiorrhiza leaf genes induced by 12-h MeJa treatment [ 53 ].…”

“…S. miltiorrhiza for 18 days. However, no studies have examined Salvia miltiorrhiza solid callus elicitation for longer time-frames, i.e., up to 60 days, and higher MeJa concentrations, up to 500 μM; such long studies have only been used to examine the influence of cytokinin, salicylic acid and auxin on tanshinone biosynthesis rate [ 37 , 38 ].…”

Improving galegine production in transformed hairy roots of Galega officinalis L. via elicitation

“…Salicylic acid (SA) is a natural phytohormone produced by plants during their normal metabolic processes (Narayani and Srivastava 2017 ) and in response to stress (Szymczyk et al 2021 ) and is involved in the signal transduction cascades of plant defense responses (Giri and Zaheer 2016 ; Ramirez-Estrada et al 2016 ). Considering the biosafety of the products, the use of these nontoxic molecules as elicitors to improve the yield of phytochemicals in vitro plant cultures has been highly recommended (Gai et al 2019 ).…”