Abstract: Artemisia vulgaris L. (mugwort) belongs to the family Asteraceae and is a tall aromatic perennial herb. Mugwort contains volatile oils, sesquiterpene lactones and flavonoids used for insecticidal, antimicrobial and antiparasitical properties. All parts of the plant are used for antihelmintic, antiseptic, antispasmodic, carminative, cholagogue, digestive, expectorant, nervine, purgative and stimulant. This study describes the effect of amino acids and growth regulators on callus induction, multiple shoot and root induction using cotyledonary explants. Murashige and Skoog medium supplemented with B5 vitamins containing 2, 4D, NAA and cysteine combination was found better response for callus induction. Maximum number of multiple shoots (95.1%) per explants after 20 days of culture, with combination of BA, TDZ and tyrosine was found better response in the medium. NAA, AgNO3 and glutamine combination produced maximum number (98.7%) of roots per explants in the medium. Plants produced from de novo regeneration on excised tissues will be useful for crop improvement through genetic engineering and cell culture techniques.