Molecular analysis of Peganum harmala L. callus to determine the gene expression of beta-carboline alkaloids harmine and harmaline

Abstract

The external signals are used as elicitors that can modify the levels of secondary metabolites production and trigger the biotechnological enforcements to improve plant production. In this study, sodium chloride NaCl was used as a stimulating factor for the production of beta-carboline alkaloids, harmine and harmaline, in Peganum harmala. The in vitro induced callus of P. harmalain was used as a source for alkaloid production in the stimulating experiments with sodium chloride. The results illustrated that 2 mg.l-1 of NaCl increased the fresh and dry weight of callus with an average of 944.30 and 72.0 mg, respectively. In a comparative analysis through Gas Chromatography (GC), high concentrations of harmine of 58.55 µg.g-1 from the root and 56.50µg.g-1 from stem callus were recorded upon treatment with 4 mg.l-1 of NaCl. Treatment with 4 mg.l-1 NaCl also showed an increased amount of harmaline concentration in both root and stem, with values of 2.72 and 2.65 µg.g-1, respectively. The analysis and calculation of gene expression by real-time PCR of RNA showed that the induced stem callus had a high gene expression with a copy number of 229,030. While in the root, the effect of higher salinity increased the percentage of alkaloids without increasing the copy number of gene expression.

Evaluation of Rosmarinus officinalis Leaves Essential Oils Activity Against Vancomycin Intermediate Staphylococcus aureus (VISA) Isolated from Baghdad Hospital Patients

“…Total RNA was extracted from treated and untreated bacteria using the TRIzol TM Reagent (Thermo Scientific, USA) [26], and checked the purity and concentration using a Quantus Fluorometer (Promega, USA). We used the SYBER green dye's, one-step quantitative RT-PCR, GoTaq® Kit 1-Step RT-qPCR System, nuclease-free water, MgCL2 and Quantifluor RNA System (Promega, USA).…”

Increasing Amounts of Secondary Metabolites and Medicinal Compounds in Callus Culture of Moringa Oleifera (Lam.) Using Abiotic Elicitors

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Influence of Water Stress on Expression of the X95950 Gene and Several Cellular Callus Components in Violet-Treated Sunflower Varieties

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