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The bacteria-free seedlings of
Przewalskia tangutica were germinated from seeds. The leaves were
used as initial explants for genetic transformation mediated by
Agrobacterium rhizogenes strain A4, with the induction of
Acetosyringone. The hairy roots were induced from the wounded sites of
the leaves at the frequency of 100%. The independently transformed and
fast growing hairy roots were subcultured in MS medium to establish the
hairy root culture lines. Genomic DNA PCR analysis was applied to detect
the presence of the rooting genes, including rolB and rolC
that had integrated into the genome of Przewalskia tangutica.
After 5-week culture in MS liquid medium, the hairy roots produced
scopolamine and hyoscyamine at higher levels by comparison to the
wild-type roots. The best scopolamine-production line produced 0.68 mg/g
DW scopolamine, and the best hyoscyamine-production line produced 1.13
mg/g DW hyoscyamine. It was the first time that hairy root cultures of
P. tangutica were established to produce tropane alkaloids.
Key words:
Przewalskia tangutica, hairy root, scopolamine, hyoscyamine. |
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