Document Type : Research Paper
Background and Objectives
Traditional propagation method of pomegranate by hard wood and soft wood cuttings does not ensure disease-free and healthy plants and it depends on the season. Hence, developing an efficient in vitro technique for the propagation of pomegranate is of great importance. On the other hand, in vitro propagation of pomegranate is also an essential step in the success of regeneration of transgenic lines and determines the effectiveness of a transformation protocol. Therefore, this study was performed to develop an efficient in vitro propagation method for ‘Malas Yazdi’ and ‘Rabab’, two Iranian leading pomegranate cultivars.
Materials and methods
For proliferation stage, WPM media supplemented with different concentrations (2.2, 4.4, 8.8 and 17.6 μM) of benzyladenine (BA) along with 0.54 μM NAA was used. To determine the effect of silver nitrate on in vitro propagation, explants were transferred to WPM medium containing 8.8 μM BA and 0.54 μM NAA supplemented with various levels of AgNO3 (5–35 μM).
For both cultivars, the best concentration of benzyladenine (BA) was 8.8 μM resulting in the highest shoot length (3.91 cm), leaf number (10.60) and node number (4.30). Adding 25 μM silver nitrate (AgNO3) to this medium significantly enhanced shoot number (4.90), shoot length (4.24), leaf number (12.30) and node number (6.10). Half-strength WPM medium supplemented with 5.4 μM NAA was most effective for rooting of shoots. Rooted plantlets were successfully acclimatized and transferred into soil. The micropropagated plants were morphologically uniform and exhibited similar growth characteristics and vegetative morphology to the mother plants.
The result of this research may be used as a step for breeding of pomegranate using genetic engineering approaches.
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