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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Keywords
Embryogenic callus , Hyoscyamus niger , Hyoscyamine