Micropropagation of Anacyclus pyrethrum and chemical profiling of the regenerated plants for pellitorine, the active principle

Abstract

The paper reports the development of a protocol for large-scale micropropagation of Anacyclus pyrethrum, an invaluable medicinal plant, and the presence of pellitorine, the active principle, in leaves and roots of regenerated plants. Three explants—cotyledonary nodes, hypocotyls and cotyledons—were compared for their shoot development/differentiation ability on Murashige and Skoog’s (MS, Physiol Plant 15:473–497, 1962) medium supplemented with 1–10 µM of 6-benzylaminopurine (BAP) or kinetin (Kn). The best response (8.88 ± 0.28 shoots per explant) was elicited from the cotyledonary nodal explants on MS + 2.5 µM Kn. The in vitro regenerated shoots continued to multiply on being sub-cultured on the same medium. Elongated shoots were cultured on MS medium augmented with different concentrations (0.2–15 µM) of 1-naphthalene acetic acid (NAA), indole-3-acetic acid (IAA) or indole-3-butyric acid (IBA) for induction of roots. The media augmented with 5 or 10 µM IAA or 5 µM NAA were the most effective for the initiation and proliferation of roots. The plantlets were successfully transferred to the following three potting mixtures: garden soil, vermiculite and garden soil (1:1), vermiculite, garden soil and perlite (1:1:1). Among these, the maximum number (75 %) of the transferred plants survived in the garden soil. The high performance liquid chromatographic analyses of roots and leaves of the regenerated plants as well as those from the native habitat revealed the presence of pellitorine in all, with the content being higher in the roots than the corresponding leaves. The developed protocol would be of use for production of A. pyrethrum plants throughout the year, and thus could become a perennial source of the herb and its active principle.