Withania coagulans (Stocks) Dunal (Solanaceae) is a critically endangered medicinal plant known for its multiple
medicinal properties. We report here an efficient and high frequency plantlet regeneration system through direct
organogenesis in W. coagulans using in vitro-derived leaves as explants. On Murashige and Skoog's (MS) medium
containing 4.44 μM 6-benzylaminopurine (BAP) 73.7 ± 4.3% explants responded and produced 11.4 ± 0.9 shoot
buds per explant. The regenerated shoot buds were elongated (6.7 ± 0.22 cm) on MS medium with 1.11 μM BAP
and 0.57 μM indole-3-acetic acid (IAA). The elongated shoots were rooted both in vitro and ex vitro. The regenerates
were acclimatized by slow and gradual exposure to different regimes of temperature and relative humidity.
The leaf-regenerates were assessed for genetic stability using RAPD and ISSR molecular markers and confirmed
true-to-type. The in vitro regeneration system developed would be useful for genetic restoration program as
the overexploitation and reproductive failure forced W. coagulans towards the verge of complete extinction.
The regeneration method would also be useful for genetic transformation for genetic improvement and
improved phyto-pharming.
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