Cloning of desert shrub Leptadenia pyrotechnica from mature shoots and their genetic stability analyses by RAPD, ISSR and ISJ markers

Leptadenia pyrotechnica (Forsk.) Decne. (Family: Asclepiadaceae) is a multipurpose shrub of arid environment. Micropropagation protocol of L. pyrotechnica was developed from mature nodal shoots. Explants were surface sterilized with 0.1% (w/v) aqueous solution of HgCl2 for 3-4 min and inoculatedion MS medium containing various concentrations of 6-benzylaminopurine (BAP) and kinetin (KIN) for activation of axillary shoot buds. Light green shoots were more responsive than dark green shoots on 5.0 mu M of BAP in comparison to KIN. Shoots were rooted in vitro on half strength MS medium containing 2.0 mu M indole butyric acid (IBA). Cultures were maintained at 28 +/- 2 degrees C, 60% RH, and 10 h photoperiods of 35-45 mu mol M-2 s(-1) (SFP) light intensity. More than 80% of in vitro regenerated plantlets were acclimatized under greenhouse conditions. The in vitro produced shoots were screened for genetic stability using RAPD, ISSR and ISJ markers. The in vitro regenerated shoots were found genetically "true to type" of the mother plant. The present protocol is an efficient and reliable mean of cloning and mass propagation of L. pyrotechnica for large-scale plantation of selected elite plants in arid environment.