A protocol for the production of high frequency multiple shoots using seedling apices with cotyledonary node of six Indian cultivars of cotton (Gossypium hirsutum L.) has been developed. An average of 7 10 shoots per explant were produced directly from the shoot apices excised from 10 day old seedlings on MS medium containing BAP (8.87?M) along with adenine sulphate (81.43 ?M) or kinetin (4.67 ?M). Rooting of isolated shoots was achieved on MS medium containing NAA (0.054?M). Rooted plantlets exhibited 95% survival during hardening in the greenhouse and regenerated plants grew to maturity with normal flowering and boll set.