Callus induction and multiple shoot proliferation from nodal explants of Mansonia altissima: confirmation of genetic stability using ISSR and RAPD markers

This study aimed to investigate indirect micropropagation of Mansonia altissima using nodal explant and confirmation of genetic stability of the plantlets in order to establish the protocol for callus induction, mass propagation, and true to type clones. Murashige and Skoog (MS) medium supplemented with 1.0, 2.0, 4.0, 8.0, and 10.0 μM IAA (indole-3-acetic acid) or 1.0, 2.0, 4.0, 8.0, and 10.0 μM IBA (indole-3-butyric acid) and 50.0 mg·L−1l-ascorbic acid was used for callus induction. The callus was subcultured on Murashige and Skoog (MS) medium containing BAP (6-benzylaminopurine) (μM) or TDZ (thidiazuron) (μM) and KIN (kinetin) (μM). The induced shoots were cultured on different strength MS medium, activated charcoal, and indole-3-butyric acid (μM) for root induction. The plantlets were acclimatized to the environment using four different types of potting media. Polymerase chain reaction amplification of the wild and indirect micropropagated M. altissima was done using 6 inter simple sequence repeats and 15 random amplified polymorphic DNA primers. Explants cultured on MS medium containing 4.0 μM IAA and 50.0 mg·L−1l-ascorbic acid had the highest percentage (95.00%) of callus formation, while 10.0 μM BAP and 0.5 μM KIN showed the highest number of shoots (17.67) and shoot height (2.03 cm) per callus. The highest number of roots (14.47) and root length (2.67 cm) was obtained in ¼ MS medium supplemented with 8.0 μM IBA and 1.0 g·L−1 activated charcoal. Moreover, plantlets were successfully acclimatized in the greenhouse. The PCR results showed true to type clones.