An efficient in vitro plant regeneration system from leaf of mature plant of Leptadenia reticulata (Jeewanti): A life giving endangered woody climber

Abstract

Leptadenia reticulata, a member of Asclepiadaceae, is a pharmaceutically important plant and has a number of medicinal properties. We describe here first time an efficient in vitro regeneration protocol using leaf of mature plant as explant. Callus was induced from cut ends of leaf inoculated on MS medium containing 1.0mgl−1 each of 2,4-D and BAP+additives. MS medium having a concentration of 2,4-D (0.5mgl−1) and BAP (0.5mgl−1) was found optimum for callus proliferation. Shoot differentiation from proliferated callus was observed on MS medium containing BAP alone or in combination with IAA or NAA. The maximum number (30.70±1.70) of shoots with average length (8.62±0.32cm) was produced on MS medium having a combination of BAP (0.5mgl−1) and NAA (0.1mgl−1). The shoots (81.6%) were rooted in vitro on one-fourth strength MS medium supplemented with IBA (1.5mgl−1) and 100mgl−1 AC. For ex vitro rooting (85.0%), shoot bases were pretreated with auxin (IBA 200mgl−1) for 3min. Rooted plantlets by both methods were successfully acclimatized under the green house conditions with about 95% survival rate. The present protocol may be applicable for (i) large scale plant production (ii) germplasm conservation (iii) genetic transformation (iv) protoplast fusion and (v) superior somaclonal variants selection of this multipurpose medicinally important endangered plant.