Establishment of in vitro regeneration from petiole explants and assessment of clonal fidelity by ISSR markers in Luffa acutangula L.Roxb
Journal Title: Journal of Applied Biology & Biotechnology - Year 2016, Vol 4, Issue 3
Abstract
An improved in vitro organogenesis was achieved from petiole explants of Luffa acutangula L. Roxb, a medicinally and economically important cucurbitaceous member. Morphogenetic response of petiole explants excised from in vitro grown plants has been studied with different concentrations of auxins and cytokinins. Higher percent of callus induction was occurred on Murashige and Skoog (MS) medium supplemented with 2mg/L BAP and 0.2mg/L NAA from the cut ends of petiole explants after 4 weeks of culture. Multiple shoots (8.28±0.85 shoots per explant) were induced after 2-3 weeks from the green compact callus on MS medium with 30g/L sucrose, 2.0mg/L BAP along with 0.2mg/L IAA. The elongation of proliferated shoots was achieved on same medium supplemented additionally with 0.3 mg/L GA3. The elongated shoots (1cm) were rooted on MS medium supplemented with 0.5 mg/L IBA. Rooted plants were acclimatized in the greenhouse and subsequently established in soil with 85% survival rate. Clonal fidelity of in vitro raised plantlets was determined using ISSR marker technique. The results showed the similar banding pattern with different ISSR primers ranging from 250bp to 1000bp and indicated the absence of polymorphism in donar mother plant and as well in in vitro regenerated plants.
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