Abstract
There is a need to enhance the production of bioactive secondary metabolites and to establish new production systems, e.g., for liver-protective compounds of Silybum marianum seeds. Quantifying and identifying the produced phytochemicals, and examining their protective effects against genotoxic agents, is of great interest. This study established a protocol for the qualitative and quantitative production of hepatoprotective compounds in cotyledon-derived Silybum marianum callus through optimized supplementation of the MS medium with the growth regulators 2,4-D, benzylaminopurine, myoinositol, and asparagine. High-performance liquid chromatography (HPLC) coupled with electrospray ionisation mass spectrometry (ESI-MS) allowed for identification and quantification of the produced compounds. None of the growth medium combinations supported a detectable production of silymarin. Instead, the generated calli accumulated phenolic acids, in particular chlorogenic acid and dicaffeoylquinic acid, as revealed by HPLC and mass spectrometric analysis. 4-Nitro-o-phenylenediamine (NPD) was employed in the AMES-test with Salmonella typhimurium strain TA98 because it is a potent mutagen for this strain. Results revealed that callus extract had a high anti-genotoxic activity with respect to standard silymarin but more evident with respect seed extract. The callus produced chlorogenic acid and dicaffeoylquinic acid, which revealed higher bioactivity than silymarin. Both compounds were not formed or could not be detected in the seeds of Silybum marianum Egyptian ecotype.
Highlights
Results revealed that S. marianum callus may serve as an alternative source for liver-protective pharmacological drugs and callus-based approaches would be of significant interest for economic and ecological reasons
The nature of the compounds, extracted from callus and seeds as well as standard silymarin, was identified by High-performance liquid chromatography (HPLC) separation followed by mass spectrometry
The results revealed that none of the callus growth regulator combination supported production and transformation of flavonoids to silymarin fractions
Summary
Gaertn (Asteraceae) is an annual to biennial herb originating from the Mediterranean and North African regions [1]. It grows wild throughout Europe, North Africa, the Americas, and Australia [2]. The flower heads of the plant are mostly purple or white. S. marianum plants flower from June to August in the Northern or December to February in the Southern Hemisphere. S. marianum fruits vary from white to nearly black [3]. The medicinal part of the plant is the seeds [4]
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