GC-MS estimation of β-asarone in diploid Acorus calamus L. and its callus-mediated regeneration

Acorus calamus L. (Sweet flag) is an important aromatic and medicinal plant used worldwide.Diploid, triploid, tetraploid and hexaploid cytotypes occur in nature and with an increase in the ploidy level, the content of -asarone increases.A high concentration of -asarone is reported as toxic and causes mutation, chromosomal anomaly and cancers.Our objective was to assess -asarone level in diploid cytotype and its callus-mediated regeneration.In this study, we investigated the -asarone concentration by gas chromatography-mass spectrometry (GC-MS).We further attempted to develop a highly effective as well as consistent callus-mediated large scale production protocol.It was found to contain low concentration of -asarone (0.67%).Optimum frequency induction of callus was carried out from shoot axillary buds which was cultured on Murashige and Skoog (MS) medium supplemented with 2 mg/l 2,4dichlorophenoxyacetic acid.MS medium fortified with 0.5 mg/l a-naphthaleneacetic acid combined with 2 mg/l 6-benzylaminopurine gave the highest regeneration (90%) results with maximum number of micro-shoots which could grow into independent plantlets.It can be inferred that such a protocol would ensure the availability of elite planting materials of diploid A. calamus for large-scale drug manufacturing operations.