High frequency plant regeneration and detection of genetic variation among micropropagated plants of Datura metel L.

摘要

High frequency plantlet regeneration was achieved from shoot tip explants of Datura metel cultured on MS medium supplemented with 6-benzylaminopurine (BAP) (2.2 micro M)+(IAA 2.8 micro M). Effect of other growth regulators including BAP, kinetin (Kn), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), alpha -naphthaleneacetic acid (NAA) and phenylacetic acid (PAA) on in vitro morphogenesis was also investigated. The highest regeneration response was observed on medium containing 2.2 micro M BAP+2.8 micro M IAA where approximately 93% of the cultures responded with an average shoot number of 24.5+or-0.7 per explant in 6 weeks time. The best elongation response occurred on MS medium with BAP (0.4 micro M)+PAA (0.7 micro M)+GA₃ (0.8 micro M) where 93% of shoots attained the average height of 5.3+or-0.3 cm. Rooting was best achieved on medium with 4.9 micro M IBA. The plantlets with well-developed shoot and root systems were acclimatized and successfully established in pots containing garden soil and organic manure. Examination of the genetic fidelity of the micropropagated plants was done by the randomly amplified polymorphic DNA (RAPD) method with 35 primers, out of which 28 could successfully generate the bands. A total of 1274 scorable bands were amplified. Among them, 169 were polymorphic, being 13% of the total bands.