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  • Author: Małgorzata Kikowska x
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Open access

Małgorzata Kikowska, Barbara Thiem, Elwira Sliwinska, Monika Rewers, Mariusz Kowalczyk, Anna Stochmal and Jolanta Długaszewska

Abstract

An efficient micropropagation protocol for production of genetically uniform clones of Eryngium campestre L. was developed. To determine the effect of nutritional and hormonal factors on shoot and root development and bioactive compounds production, three variants of media differing in the content of macro- and micronutrients, as well as plant growth regulators of various types and concentrations were tested. The highest regeneration (100%), with over 13 shoots per explant, was induced on Murashige and Skoog (MS) medium with 1.0 mg l−1 benzyladenine (BA) and 0.1 mg l−1 indole-3-acetic acid (IAA). The in vitro derived shoots multiplied through axillary bud formation were rooted and transferred to an experimental plot with 78% frequency of survival. Flow cytometry showed no variation in nuclear DNA between the seedlings and micropropagated plants. Preliminary thin layer chromatography (TLC) analysis indicated that phenolic acids, saponins, flavonoids and acetylenes were present in plant biomass. Ultra high performance liquid chromatography (UHPLC) analysis revealed that shoots and roots from in vitro derived plants and root cultures maintained the ability to produce rosmarinic acid (RA), rosmarinic acid hexoside (RA-HEX) and chlorogenic acid (CGA). The highest phenolic acid content was detected in roots of in vitro regenerated plants. The extract from those roots expressed the highest inhibitory effect against bacteria Staphylococcus aureus, as well as dermatophytes Trichophyton mentagrophytes and T. rubrum.

Open access

Małgorzata Kikowska, Agata Włodarczyk, Anna Stochmal, Jerzy Żuchowski and Barbara Thiem

Summary

Introduction: Callus and cell suspension cultures are widely applied in investigation of production of high-value secondary metabolites, which may be used as cosmeceuticals, nutraceuticals and pharmaceuticals. Plant cell cultures are promising alternative to intact plant sources for the production of plant-derived drugs of industrial importance.

Objective: The aim of the study was to (i) initiate the cell suspension culture of Chaenomeles japonica from homogenous and uniform callus, (ii) stabilize the selected line and (iii) verify its ability to produce the desired groups of secondary metabolites – pentacyclic triterpenoids and polyphenols.

Methods: To establish a cell suspension culture, stabilized and homogeneous callus was selected. Cell cultures were systematically passaged every 2 weeks to fresh liquid medium with the same composition. Biomass from cultures at the growth phase and stationary phase was designated for phytochemical research. UHPLC-DAD-MS analyzes were performed. At the same time, their macroscopic and microscopic observations were carried out.

Results: Cells of suspension culture line A2 were characterized by the intense divisions. Cell culture extracts (both from the growth phase and stationary phase) contained pentacyclic triterpenoids. In addition, phe-nolic compounds (chlorogenic acid and proanthocyanidins type B) and in a small amount also epicatechin are present in the extract of the cells harvested from the growth phase. In the present studies, three pentacyclic triterpenoids were detected and quantified in the extracts of cell suspensions and callus line A2. Ursolic and oleanolic acids were the main triterpenoids in the studied extracts. The cell suspension culture from the growth phase exhibited the highest content of ursolic, oleanolic, and betulinic acid (separately and together).

Conclusion: The cell suspension culture of Chaenomeles japonica is a promising source of pentacyclic triterpenoids.

Open access

Małgorzata Kikowska, Jolanta Długaszewska, Marcelina Maria Kubicka, Izabela Kędziora, Jaromir Budzianowski and Barbara Thiem

Summary

Introduction: Due to increasing resistance against antibiotics and antifungal agents, crude plant extracts, fractions, and isolated pure compounds became a new interest as antimicrobial agents.

Objectives: The antimicrobial activity of methanolic extracts and fractions of Eryngium planum L., E. campestre L., and E. maritimum L. was evaluated against selected bacteria, yeast and mould, and compared in tested Eryngium species and in their organs.

Methods: The antimicrobial activity was studied with use of broth microdilution method. The antibacterial (Staphylococcus aureus, Pseudomonas aeruginosa) and antifungal (Candida albicans, Aspergillus niger) activity of selected extracts and fractions compared with the reference substance was expressed by Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal/Fungicidal Concentration (MBC/MFC). The extract and fraction compounds were identified on the basis of TLC examination.

Results: The saponin-phenolic acid fractions of E. maritimum and E. planum and a saponin fraction of E. planum showed the highest activity against S. aureus (MIC = 1–2.5 mg·ml−1). The growth of C. albicans was inhibited by methanolic extract of E. planum cell suspension culture (MIC = 7.8 mg·ml−1).

Conclusion: The antimicrobial activity depends on the Eryngium species, tested biomass, and microorganism.