The Establishment of embryogenic callus culture of hyoscyamus niger and the detection of hyoscyamine in the culture

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Date
2008-05
Authors
Saidon, Nhawal Aminie
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Abstract
Embryogenic callus of Hyoscyamus niger could be induced from the leaf, petiole and root explants however the best embryogenic callus induction was obtained from the leaf explants cultured on MS medium supplemented with 6.0 mg/L picloram after four weeks of culture. The embryogenic callus could undergo maturation stage on MS supplemented with 1.0 mg/L BA followed by one week culture on MS basic medium. The addition of casein hydrolylasate in the MS + 1.0 mg/L BA medium generally induced higher biomass of embryogenic callus and more globular and torpedo shape embryos. However, the mature somatic embryos failed to germinate in the germination medium, MS supplemented with 0 - 10.0 mg/L BA. Embryogenic cells of H. niger could be propagated by culturing the friable embroygenic callus in liquid MS medium supplemented with 6.0 mg/L picloram and subculturing was carried out every 12 days. Transferring the embryogenic cells into liquid MS medium only produce root without showing any sign of maturation. Preliminary study showed that direct embryogenesis could be induced from the leaf and petiole explants cultured on MS medium supplemented with BA (2.0 - 8.0 mg/L) while only petiole explants showed direct embryogenesis formation on MS medium supplemented with 6.0 mg/L BA and 0.5 mg/L NAA. Histological study confirmed the formation of both indirect and direct embryogenesis of H. niger. GCMS analysis revealed the presence of hyoscyamine in the H. niger embryogenic callus induced on MS supplemented with 1.0 mg/L BA after culturing on MS for one week and embryogenic callus and somatic embryos cultured on MS + 500 mg/L casem hydrolysate.
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Embryogenic callus , Hyoscyamus niger , Hyoscyamine
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