Abstract
The present investigation was carried out to for developing a competent protocol for micropropagation of commercial plum cultivar Frontier, through control of shoot tip necrosis (STN) observed during in vitro shoot multiplication. Apical and axillary buds were collected from elite high yielding tree of Frontier and inoculated on Murashige and Skoog (MS) medium fortified with phytohormones at different concentrations. MS medium containing 0.5 µM BA, 0.5 µM Kin and 0.2 µM IBA resulted in highest shoot bud establishment (58.95%) after 4 weeks interval. However, all the shoots showed necrosis and died after 1st subculture. Necrosis appeared with yellowing of shoot tips within 7 days of first subculture and gradually increased downwards resulting in yellowing of leaves, followed by death of the shoots. Among various reported methods tested to control STN, combination of Fructose (0.50 mM) and Calcium chloride (1.0 mM) proved 100% effective. Highest in vitro shoot multiplication (fivefold) was recorded on medium enriched with 2.0 µM BA, 0.2 µM GA3, 0.2 µM IBA, 0.50 mM Fructose and 1.0 mM Calcium chloride. For rooting, two step rooting procedure was followed which resulted in 85.00% rooting when microshoots were given a dark treatment by dipping in ½ MS broth supplemented with 2.5 µM IBA for 48 h before transferring to semisolid ½ MS basal medium. Rooted plantlets when transferred in vivo for acclimatization, showed 70–80% survival after 10 weeks and no mortality on field transfer. Molecular analysis of the plants using ISSR primers proved 100% similarity between them and mother tree.
Published Version
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