A high frequency secondary somatic embryogenesis and plant regeneration have been achieved in Calliandra tweedii (Benth.) through internodal segments excised from 5-year-old shrub on Murashige and Skoog (MS) medium enriched with different concentrations of 2-isopentenyl adenine (2iP). Somatic embryos were differentiated directly from cut ends, epidermal crevices as well as indirectly through callus. Such primary embryos when subcultured on MS medium augmented with 2-isopentenyl adenine (2iP), N6 benzyladenine (BA), 2,4-dichlorophenoxy acetic acid (2,4-D), α-naphthaleneacetic acid (NAA) in different concentrations (0.1, 0.5, 1, 1.5, 5, 10, and 20μM) individually, redifferentiated numerous secondary somatic embryos of different stages, i.e., globular, heart-shaped, torpedo and cotyledonary types within 6 weeks of culture. The somatic embryos originated either from radicular or plumular parts or from all over the surfaces. Of the aforesaid growth regulators, 1μM 2iP proved to be the best for recurrent production of an average of 19.44±0.52 cotyledonary somatic embryos per culture within 6 weeks. The embryogenic clumps (100mg per culture tube) comprising the pre cotyledonary stages were transferred to different concentrations (1, 2.5, 5, 7, 10, 20, and 50μM) of abscisic acid (ABA) for their maturation and preventing precocious germination. ABA at 5μM proved optimum for developing an average of 15.87±0.31 normal cotyledonary embryos per embryogenic clumps within 4 weeks. These bipolar mature embryos after transfer to half-strength MS medium germinated into plantlets. The genetic uniformity of the emblings was established employing 14 inter simple sequence repeat (ISSR) marker where 96.1% of the bands generated were monomorphic and similar with those of stock plant with only 2% variation among the plantlets.