The active substances in chamomile (Matricaria recutita L.) belong to chemically different structural types. The largest group of medically important compounds forming the essential oils are primarily chamazulene, (-)-alpha-bisabolol, bisabololoxides, bisabolonoxide A, trans-beta-farnesene, alpha-farnesene, spathulenol and the cis/trans-en-in-dicycloethers. Flavonoids, coumarins, mucilages, mono- and oligosaccharides also have pharmacological effects. We studied the production of essential oils in genetically transformed cultures. Sterile juvenile chamomile plants were infected with A4-Y strains of Agrobacterium rhizogenes. They are known plant pathogens and are capable of inducing so-called hairy roots. The transfer DNA segment of the Ri-virulence plasmid of A. rhizogenes becomes integrated in the genome of the plant cells. The isolated hairy roots grow rapidly on hormone-free media. In order to obtain bacteria-free media, we cultured the transformed roots on Murashige-Skoog (MS) medium supplemented with carbenicillin (800 mg/l). To study the production of essential oils, the clones were propagated on liquid and solid MS and Gamborg (B5) media, respectively. According to gas chromatography, the composition of the essential oil of hairy root cultures on different media was found to be similar, but differing in proportion. The main component of the essential oil which was identified by gas chromatography and mass spectrometry was trans-beta-farnesene, as in the intact roots.
We studied the essential oil production of cultivated (BK-2, Degumil) and wild chamomile populations of 4 typical chamomile-rich regions of Hungary. We examined the essential oil composition of flowers, herbs (stem plus leaves) and roots using GC and GC/MS methods. Among cultivated species, the Hungarian BK-2 contains more chamazulene in its essential oil than the German Degumil type, which is mainly cultivated for its α-bisabolol. Both components have important antiinflammatory activities. Wild populations can be easily distinguished from cultivated ones by their high amount of bisaboloides, particularly the flower of Szabadkígyós wild type, which contained on average 48 % of the biologically active (-)-α-bisabolol. The regional wild chamomile samples mentioned above have already been examined previously in our Institute. We found it interesting to compare the content of biologically active components, at the same conditions, of presently promising populations with the results obtained 20 years ago from the same species. While the content of the essential oil of rural Szabadkígyós wild type remained unchanged, there was a trend of the essential oil components towards the therapeutically important compounds. The amount of (-)-α-bisabolol in Szabadkígyós mounted up to a 3-fold increase and we measured a doublefold increase of chamazulene content in BK-2 compared with samples 20 years ago. We can conclude that although a change was observed in the essential oil content and also in the proportion of different components, the fundamental characteristics of the oils remained the same. To keep the genom of Szabadkígyós wild type having high (-)-α-bisabolol content, we used biotechnological methods. The sterile roots of organised culture contained also β-eudesmol, wich was firstly identified from the intact roots by us. Our gas chromatographical and mass-spectroscopical studies showed that sterile chamomile cultures generated the most important terpenoid and polyin compounds characteristic of the mother plant. We identified berkheyaradulene, α-selinene, geranyl-isovalerate and cedrol as new components in these sterile cultures.
We identified alpha-selinene, as a new component in these hairy roots. We studied the growth rate of A4-Y clone on the cited media, containing MgSO4 concentrations: 0; 185; 370 and 740 mg/l. The cultures grew most in medium containing 740 mg/l of MgSO4. Essential oil content was compared from hairy root cultures of different Mg containing media and measured by GC and GC-MS methods. Mg has a similar effect on hairy roots as on organized cultures.
Gas chromatography (GC) and GC-mass spectrometry are used to identify a new sesqiterpene, beta-eudesmol, which seems to be a characteristic essential oil component of the intact and in vitro organized root of chamomile [Chamomilla recutita (L.) Rauschert]. It is identified on three types of stationary phases by GC. The confirmation of identity is carried out by comparison of mass spectra with those reported in the literature and measured from a reference compound. The percentage evaluation of the oil component is made by area normalization, on the basis of three parallel measurements. Among the cultivated and wild chamomile species examined, the wild species from the areas of Szeghalom contain the highest quantity of beta-eudesmol (9.25% in the total essential oil).
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