Abstract
The aim of this study was to assay by NMR the metabolites which contribute to betulin production. 8-day-old suspended birch (Betula platyphylla) cells were treated by sodium nitroprusside (SNP) treatment, an NO donor, and 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (cPTIO), an NO-specific scavenger. The results showed that betulin production was increased by five times after SNP treatment, similar with that of the control under cPTIO treatment. Forty one metabolites were detected after SNP treatment or cPTIO treatment. Among them, 10 were found to significantly contribute to the differences observed between controls and treated cell culture samples. To validate the contribution of the above 10 metabolites to betulin production, myo-inositol, fructose and phenylalanine based on correlation analysis between the content of 12 metabolites and betulin were used to feed birch suspension cell cultures under SNP treatment. Exogenous feeding of fructose or phenylalanine further enhanced the betulin production under SNP treatment, but myo-inositol had the opposite result.
Highlights
In vitro experimentation on plant tissue cultures gives a wide range of secondary metabolites that are used as therapeutics, flavors, fragrances, colors, and food additives [1,2]
Figure production of the birch suspension cellscells induced by sodium nitroprusside (SNP)
The results showed that SNP treatment changed metabolites and betulin under 1 mmol/L SNP
Summary
In vitro experimentation on plant tissue cultures gives a wide range of secondary metabolites that are used as therapeutics, flavors, fragrances, colors, and food additives [1,2]. For increasing the entire biomass and production of high amounts of secondary metabolites in plant cell cultures, the use of biotic and abiotic elicitors has been recognised as one of the most effective strategies [3,4]. Some such pharmaceutical secondary metabolites, the triterpenoids betulin and oleanolic acid, can be extracted from the bark of Betula platyphylla Suk. Triterpenoids are excellent drug candidates, with antiviral, antibacterial, antitumor, and anti-AIDS activity [5,6,7]. The biosynthesis of NO and plant secondary metabolites was reported to protect plants from attack by insect, herbivores, and pathogens, or to survive under abiotic stresses, and NO plays important roles in the accumulation of secondary metabolites in plants [12], so a better understanding of the role of NO in the biosynthesis of such secondary metabolites is very important for optimizing the Molecules 2017, 22, 1035; doi:10.3390/molecules22071035 www.mdpi.com/journal/molecules commercial production productionof of those pharmaceutically significant secondary metabolites
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