2012•JoLS Journal of Life SciencesRequires access
Alkaloids Production from Callus of Hyoscyamus niger L. in Vitro
Abedaljasim M. Jasim Al-Jibouri, Khulood Whaybed Al-samarraei, Ashwaq Shanan, Duhaa Muasar Mageed, Abdal-Jabbar Abass Ali
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Abstract
Callus cultures of Hyoscyamus niger L. were initiated from leaf segments cultured on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L Benzyl Adenine (BA) and 0, 1, 2 and 3 mg/L Naphthalene acetic acid (NAA). Half of cultures were incubated under light of 16 hr/day, while the other half was incubated under complete darkness. The incubation temperature was 25 ± 1 °C in both incubation conditions. The fresh and dry weight of the produced callus was obtained after six weeks of incubation. Callus produced were recultured on medium that gave the highest production of callus. Constant weight (300 mg) of callus was cultured in each of these medium supplemented with abiotic elicitor of 50 g/L sucrose, 200 mg/L NaCl, 50 or 100 mg/L proline and 2 mg/L BA each one added separately and incubated under complete darkness. The fresh and dry weights of callus were measured after six weeks. HPLC was used to determine the tropane alkaloids (Hyoscyamine and Scopolamine). The results showed that the significant highest average of fresh and dry weight of callus (112 and 89.6 mg) achieved using the medium contained 0.5 mg/L BA and 2 mg/L NAA under dark condition. The amount of fresh and dry weight of callus produced under dark condition was significantly higher than that produced under light condition, with increase in percentage of 51.3 and 37.62% respectively. The addition of abiotic elicitors caused reduction in both fresh and dry weight of callus, therefore the highest fresh weight average was 1,727 mg using 100 mg/L proline. The results indicated that addition of 50 or 100 mg/L proline led to increase in Hyoscyamine concentration of 58.03 and 21.37% respectively compared with the control. While other abiotic elicitors were found to cause reduction in Hyoscyamine concentration. Percentage of Scopolamine concentration were increased to 129.03, 166.94, 205.51 and 149.20% after addition of sucrose (50 g/L), NaCl (200 mg/L) and proline (50 or 100 mg/L) respectively compared with the control.