Evaluation of genetic fidelity among micropropagated plants of Salvadora persica L. using DNA-based markers

Salvadora persica L. (family Salvadoraceae) is a highly threatened medicinally and economically important arid zone plant species. With ever increasing demand, there is a pressing need for conservation and mass multiplication of this plant through biotechnological approaches. A large number of complete plantlets were obtained by direct regeneration from the nodal explants on Murashige and Skoog (MS) medium supplemented with 2.0 mg/l 6-benzylaminopurine (BAP) + 1.0 mg/l indole-3-acetic acid (IAA) containing 30 g/l sucrose and 8 g/l agar at 16 h photoperiod. A high frequency of in vitro rooting was obtained on half strength MS medium supplemented with 3.0 mg/l indole-3-butyric acid (IBA). The micropropagated plantlets were hardened using sterilized soil and sand mixture (3:1) and successfully transferred under field conditions. The genetic stability of micropropagated clones was evaluated using random amplified polymorphic DNA (RAPD) analysis. During the study a total of 43 RAPD primers were screened, out of which 8 primers produced a total of 37 clear, distinct and reproducible amplicons. The amplified products of all the regenerated plants were monomorphic and were similar to the mother plant, thus confirming the genetic homogeneity of tissue culture raised plants. To the best of our knowledge, it is the first report on the assessment of the genetic fidelity in micropropagated plants of S. persica, which could be successfully applied as one of the safest modes for the production of true-to-type plants for the germplasm conservation.