In vitro regeneration of gamma irradiated callus of artemisia annua and evaluation of increase artemisinin content by HPLC analysis

Abstract

Gamma radiations are the efficient ionizing radiations which have potential to induce mutations in plants They also modify physiological characteristics of plant to create new mutants for large scale production of commercially important secondary metabolites In present study the effect of various low doses gamma irradiations and Gy on growth regeneration and elicitation of Artemisinin content from Artemisia annua was studied Callus culture was established on Murashige and Skoog medium supplemented with phytohormones Napthalein acetic acid and N benzylaminopurine mgL minus Effect of gamma doses on callus growth and elicitation of artemisinin content were investigated from M T M T M T M T and M T generation of in vitro callus culture In current study it was found that low dose of gamma irradiations are effective strategy to enhance the yield of artemisinin in A annua The radiation sensitivity test was performed which is based on the reduction in survival rate of callus culture after treatment with gamma irradiation at different doses from to Gy which showed the negligible effect till Gy but at and Gy it inhibited the callus growth and callus regeneration The detection and quantification of artemisinin content was performed by HPLC analysis The mobile phase for artemisinin elution was acetonitrile water v v at a flow rate of ml min minus and UV detection at nm Artemisinin concentration of unknown sample was calculated by using the Y equation y x R sup generated from standard curve The retention time of artemisinin in above solvent system was observed at min The higher accumulation of artemisinin content was observed in Gy dose micro g gm dry callus weight which is tenfold higher than non treated micro g gm dry callus weight The present study confirms that gamma radiation induced mutational changes in callus culture and it enhanced the artemisinin content