In-vitro propagation of the multipurpose Egyptian medicinal plant Pimpinella anisum

Background and objective Plant tissue culture technology offers a solution for meeting the increasing commercial demand for medicinally important plants, especially cross-pollinating species, whose genetic heterogeneity presents difficulties when using traditional propagation methods. Herein, we describe an effective, rapid, and simple protocol for the micropropagation of anise (Pimpinella anisum). Materials and methods We investigated the effect of the type of explant and the type and concentration of plant growth regulator, either individually or in combination, on plant micropropagation. Results and conclusion Although multiple shoot formation rate was higher for nodal than for shoot tip explants, there was no significant difference in rooting response between shoots arising from either of them. Maximum shoot response, number of shoots per explant, and shoot length were observed in nodal explants grown on Murashige and Skoog medium supplemented with 5 mu mol/l 6-benzylaminopurine, 1 mu mol/l kinetin, and 0.5 mu mol/l naphthalene acetic acid. The most effective medium for root regeneration contained 3 mu mol/l of either indole-3-butyric acid or naphthalene acetic acid. Interestingly, there was no evidence for hyperhydricity, which is commonly found in cultured anise, using our method. Plantlets were successfully hardened and transferred to the greenhouse, with an 85% survival rate. This protocol provides an efficient means for large-scale production of anise, as well a basis for further research aimed at genetic improvement of this plant.