Study the Effect of Some Biotic and Abiotic Factors on Enhancement of Essential Oils and Rosmarinic Acid in Rosemary Rosmarinus officinalis L. In vitro

The yield of rosemary (Rosmarinus officinalis L.) from essential oils and rosmarinic acid were investigated. Yield of callus tissue was compared with the intact plant production. Callus was induced on leaf explants and maintained on Murashige and Skoog medium (MS) supplemented with Dichlorophenoxy acetic acid (2,4-D), Benzyl adenine (BA) and Kinetin (KIN). The highest percentage of callus induction (100%) was recorded on leaf explants when a combination of 2 mg/l 2,4-D and 0.5 mg/l BA was added to the culture medium followed by (90%) at the combination of 2 mg/l 2,4-D and 0.1 mg/l KIN. Maximum callus fresh weight was obtained in the combination of 2 mg/l 2,4-D and 0.5 mg/l BA in cultures grown under 16/8 hrs photoperiod, which reached 1780 mg, while it recorded 1442.5 mg in the combination of 2 mg/l 2,4-D and 0.1 mg/l KIN. Callus cultures were treated with biotic (bacterial, fungal and yeast extract) and abiotic elicitors (CaCl2 and gamma-rays). The quantitative and qualitative analysis of essential oils was carried out using gas chromatography-mass spectrometry (GC-MS). The most abundant contents of the rosemary leaves essential oils were, camphor (20.54%), cineole (13.40%), verbenone (12.71%), bornyl acetate (12.22%), carene (7.73%), camphene (4.32%), α- pinene (2.28%) and β-pinene (1.77%). The most presented components of leaves ethanol extract were, isocarnosol (5.85%), camphor (3.88%), ferruginol (4.53%), cineole (2.66%), verbenone (2.50%) and borneol (1.56%), while the most presented components of callus ethanol extract were, cineole (4.44%), camphor (2.51%), verbenone (2.30%), borneol (1.91%), bornyl acetate (1.66%).