Abstract
Callus initiation, shoot formation and plant regeneration were established for Artemisia spicigera, a traditional medicinal plant growing in Armenia, Middle-Anatolia and Iran, and producing valuable volatile organic compounds (VOCs) that are mostly represented by monoterpenoids. Optimal callus initiation and shoot production were obtained by culture of hypocotyl and cotyledon explants on MS medium comprising 0.5 mg L−1 naphthalene acetic acid (NAA) and 0.5 mg L−1 6-benzyladenine (BA). Consequently, the shoots were transferred onto the MS media supplemented with 1 mg L−1 of indole-3-butyric acid (IBA) or 1 mg L−1 of NAA. Both types of auxin induced root formation on the shoots and the resulting plantlets were successfully grown in pots. The production of VOCs in callus tissues and regenerated plantlets was studied by gas chromatography–mass spectrometry (GC-MS) analysis. Although the potential of undifferentiated callus to produce VOCs was very low, an increased content of bioactive volatile components was observed at the beginning of shoot primordia differentiation. Intriguingly, the volatiles obtained from in vitro plantlets showed quantitative and qualitative variation depending on the type of auxins used for the rooting process. The acquired quantities based on total ion current (TIC) showed that the regenerated plantlets using 1 mg L−1 NAA produced higher amounts of oxygenated monoterpenes such as camphor (30.29%), cis-thujone (7.07%), and 1,8-cineole (6.71%) and sesquiterpene derivatives, namely germacrene D (8.75%), bicyclogermacrene (4.0%) and spathulenol (1.49%) compared with the intact plant. According to these findings, in vitro generation of volatile organic compounds in A. spicigera depends on the developmental stages of tissues and may enhance with the formation of shoot primordia and regeneration of plantlets.
Highlights
There was no significant difference in fresh weights of the calli formed on media with altered combinations of naphthalene acetic acid (NAA)
The dry weight of calli originated from both hypocotyl and cotyledon explants in the concentration of 0.5 + 0.5 mg L−1 NAA +
These results indicated that the medium containing 0.5 mg L−1 of NAA + 0.5 mg L−1 of BA is more suitable for the establishment and growth of callus tissues of A. spicigera in comparison with other treatments (Table 1)
Summary
Plant tissue culture is an important technique applied in different areas of research for micropropogation, secondary metabolites production, and toxicological studies [1,2,3]. The application of tissue culture for studying the regeneration potential and changes in the chemical composition is very important to improve the production of natural products obtainable from valuable medicinal and aromatic plants. (Asteraceae) is a large, diverse, and widely distributed genus with more than 500 species. This genus comprises of bitter, aromatic herbs or shrubs, which are famed for the production of volatile organic compounds (VOCs) of medicinal value [5,6]
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