Abstract
Artemisia vulgaris L. produces a wide range of valuable secondary metabolites. The aim of the present study is to determine the effects of various concentrations of farnesyl diphosphate (FDP) on β-caryophyllene content in both callus and hairy root (HR) cultures regeneration from leaf explants of A. vulgaris L. Murashige and Skoog (MS) medium supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4D; 4–13 μM), α-naphthaleneacetic acid (NAA; 5–16 μM), and FDP (1 and 3 μM) was used for callus induction and HR regeneration from leaf explants of A. vulgaris L. In this study, precursor-treated (2,4D 13.5 μM + FDP 3 μM) callus displayed the highest biomass fresh weight (FW)/dry weight (DW): 46/25 g, followed by NAA 10.7 μM + FDP 3 μM with FW/DW: 50/28 g. Two different Agrobacterium rhizogenes strains (A4 and R1000) were evaluated for HR induction. The biomass of HRs induced using half-strength MS + B5 vitamins with 3 μM FDP was FW/DW: 40/20 g and FW/DW: 41/19 g, respectively. To determine β-caryophyllene accumulation, we have isolated the essential oil from FDP-treated calli and HRs and quantified β-caryophyllene using gas chromatography–mass spectrometry (GC–MS). The highest production of β-caryophyllene was noticed in HR cultures induced using A4 and R1000 strains on half-strength MS medium containing 3 μM FDP, which produced 2.92 and 2.80 mg/ml β-caryophyllene, respectively. The optimized protocol can be used commercially by scaling up the production of a β-caryophyllene compound in a short span of time.
Highlights
Artemisia vulgaris L. is an important medicinal herb that belongs to the family Asteraceae
fresh weight (FW)/DW of callus 2,4D treated without precursor was observed at FW/DW: 36/22 g, whereas in precursor-treated callus the highest FW/DW: 48/28 g were noticed at the concentrations (2,4D 13.5 μM + farnesyl diphosphate (FDP) 3 μM) followed by (2,4D 9 μM + FDP μM) fresh and dry weight was exhibited (Figure 2A)
Similar results were observed in Psoralea corylifolia with precursor treatment in cotyledonary callus, which was obtained from FW 2,601.8 mg/g (Mohammadparast et al, 2015)
Summary
Artemisia vulgaris L. is an important medicinal herb that belongs to the family Asteraceae. A. vulgaris L. has been widely used as a traditional medicine for treating a number of diseases, such as neonatal jaundice (Fok, 2002), gastric ulcers (Repetto and Llesuy, 2002), and hepatitis, as well as an analgesic agent in acupuncture therapy (Yoshikawa et al, 1996). It has been effective as antiviral (Tan et al, 1999), insect-repellant, fumigant (Wang et al, 2006), insecticide (Chantraine et al, 1998), antibacterial, anti-inflammatory (Tigno et al, 2000), sedative, and flavoring and perfumery agents (Da Silva, 2004). It has been found that some species of the plant kingdom produce sesquiterpene β-caryophyllene (De Moraes et al, 2001)
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