Indian Journal of Biotechnology Vol 5, April 2006, pp 217-222 Somatic embryo proliferation in Commiphora wightii and evidence for guggulsterone production in culture Sandeep Kumar 1 , Meeta Mathur, A K Jain and K G Ramawat* Laboratory of Bio-molecular Technology, Department of Botany, M L Sukhadia University, Udaipur 313 002, India 1 G B Pant Institute of Himalayan Environment & Development, Sikkim Unit, PO Tadong, Gangtok, Sikkim 737 102, India Received 6 December 2004; revised 6 June 2005; accepted 2 August 2005 A method for obtaining somatic embryos consistently in Commiphora wightii (Arnott.) Bhandari has been developed. Somatic embryos when sub-cultured on modified MS medium containing ABA (10 μg L -1 ) produced more embryos directly by meristematic activity of the epidermal and subepidermal cells at the hypocotyl region or also from the root tip region. Embryo proliferation was also observed on auxin-cytokinin free modified MS medium supplemented with different concen- trations of activated charcoal, abscisic acid, mannitol and sucrose. Modified MS hormone-free medium containing activated charcoal 0.5 g L -1 and sucrose 10 g L -1 was most suitable for multiplication and maturation of embryos. Embryo formation was also recorded from callus produced by earlier formed somatic embryos (indirect). Embryos obtained from various matu- ration treatments when desiccated under laminar airflow bench for 5-9 d showed maximum germination percentage. Gug- gulsterone-E and -Z contents of in vitro embryonic cultures were ¼ - and ½ -fold of that found in zygotic embryos, but sev- eral fold higher than callus cultures, therefore, hold promise for the production of bioactive guggulsterones. Keywords: Commiphora wightii, somatic embryogenesis, guggulsterones, antihyperlipidemia, callus cultures IPC Code: Int. Cl. 8 A01H4:00; A61P3/06 Introduction Commiphora wightii, a balsamiferous shrub, is an important medicinal plant which yields an oleogum- resin known as Guggul. Guggul is used extensively by Ayurvedic physicians to treat a variety of afflictions, including arthritis, inflammation, bone-fractures, obe- sity and disorders of lipid metabolism 1 . The plant has been over exploited for its gum resin and as a fuel- wood besides it’s slow growth, poor seed set has made it endangered species 2 . The gum resin contains active components guggul- sterones, which are effective in controlling high blood cholesterol and lipids 3-8 . Several reports have conclu- sively established through modern pharmacological tests the hypolipidemic and hypocholesterolemic properties of C. wightii extract. Guggulsterones lower cholesterol by acting as antagonist to farnesoid X re- ceptors, a nuclear hormone receptor that is activated by bile acid 9-10 . The biology, chemistry, biotechno- logical approaches for the amelioration of plant and mode of action of guggulsterones has already been reviewed 11,12 . Earlier, the authors reported the formation of so- matic embryos from immature zygotic embryo ex- plants of C. wightii 13 and subsequently the establish- ment of embryonic callus and formation of somatic embryos 14 . Histological studies showed formation of resin canal in somatic embryos at torpedo stage 15 . It is of interest to develop method for continuous somatic embryo formation and to examine these cultures for possibility of producing guggulsterones. In this com- munication we report embryonal proliferation and presence of gugulsterones in the cultures. Materials and Methods Immature fruits (5-6 mm) of Commiphota wightii (Arnott.) Bhandari collected from the plants growing wild in Madar near Udaipur were used to initiate cal- lus cultures. Surface sterilization, dissection and cul- ture of zygotic embryos from these fruits, medium preparation and culture conditions were same as de- scribed earlier 14 . Callus obtained from zygotic em- bryos on B5 medium 16 containing 2,4,5- trichlorophenoxy acetic acid (2,4,5-T) and kinetin was transferred and maintained on hormone-free (HF) B5 medium for 3-4 passages. Details of somatic embryo- genesis in callus cultures are reported in the previous work 14 . —————— *Author for correspondence: Tel: 91-4144-243533 extn. 210: Fax: 91-4144-243555, 238419 E-mail: cdl_aucasmb@sancharnet.in; cdl_mangrove@sancharnet.in