IN VITRO MULTIPLE SHOOT REGENERATION AND ANALYSIS OF GENETIC FIDELITY OF MENTHA PIPERITA L. | BIONAT

An efficient, rapid and reproducible plant regeneration protocol was successfully developed for Mentha piperita L. using nodal explants from naturally field grown plants. Axillary shoot bud proliferation, initiated from nodal explants was cultured on MS (Murashige and Skoog, 1962) medium supplemented with various concentration of 6-benzyladenine (BA) (0.5-3.0 mg/l), Kinetin (KN) (0.5-3.0 mg/l) and 2-isoPentenyladenine (2-iP) (0.5-3.0 mg/l). Maximum numbers of shoots (4.19 ± 0.03) with average length (3.46 ± 0.16) were induced on medium containing 1.0 mg/l BA. Shoots culture was established by repeated sub-culturing the original nodal explant on shoot multiplication medium after each harvest of newly formed shoots. Thus, 20-25 shoots were obtained in 3 months from a single nodal explant. Rooting of shoots was achieved on half strength MS medium supplemented with 1.5 mg/l Indole-3-butyric acid (IBA) and 2% sucrose. Welldeveloped complete plantlets were transferred to plastic pots containing a mixture of (1:1) soil and vermiculite showed 81.13% survived. The hardened plants were evaluated for their genetic fidelity through RAPD markers. Because, genetic uniformity of in vitro raised plants as also a prerequisite for production of quality planting material of Mentha piperita L. This protocol can be used for commercial propagation purposes and to initiate future genetic improvement studies.

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