J. Acad. Indus. Res. Vol. 1(5) October 2012 281 ©Youth Education and Research Trust (YERT) Nomila Merlin et al., 2012 ISSN: 2278-5213 Taxol production by endophytic Fusarium solani LCPANCF01 from Tylophora indica J. Nomila Merlin 1 , I.V.S. Nimal Christhudas 2 , P. Praveen Kumar 2 , M. Kumar 3 and P. Agastian 2 1 Dept.of Biotechnology, Bharathiar University, Coimbatore-641046, TN, India 2 Dept. of Plant Biology and Biotechnology, Loyola College, Chennai-600034, TN, India 3 Dept. of Plant Biology and Plant Biotechnology, Madras Christian College, Tambaram, Chennai-600059, TN, India agastian@loyolacollege.edu; +91 9444433117 _____________________________________________________________________________________________ Abstract Twenty five endophytic fungal isolates were obtained from the roots of Tylophora indica (Burm. f) and screened for the presence of an anticancer drug, taxol. Among the 25 isolates, Fusarium solani LCPANCF01 was identified based on the micro morphology, cultural characteristics and sequence analysis using internal transcribed spacer (ITS1 and ITS4). Fusarium solani LCPANCF01 strain was grown in M1D liquid medium for 21 d and extracted with dichloromethane. The presence of taxol was confirmed by TLC, HPLC, UV, IR, and ESI-MS spectroscopy analysis by comparing with the standard drug. Keywords: Tylophora indica, Fusarium solani, dichloromethane, taxol, anticancer drug. Introduction Taxol is isolated from the barks of pacific yew tree (Taxus brevifolia) (George et al., 1994; Dewick, 2009) and the most important antimitotic agent which is active against lung, ovarian, breast, head-neck cancer and advanced forms of Kaposi's sarcoma. Taxol inhibits cell proliferation by binding to the β-subunit of the tubulin heterodimers, thus promoting its polymerization (Kovacs et al., 2007). However, a complete course of treatment per patient may requires 2 g of taxol administered several times over many months. To obtain 1 kg of taxol, it requires 3000 yew trees (10,000 kg of bark) and current demand of taxol is 250 kg per annum (Dewick, 2009). This insufficiency of taxol and environmental issue of harvesting from trees demands researcher to discover the alternate technique for the production of taxol. Taxol is now obtained by several methods, e.g. total chemical synthesis (Nicolaou et al., 1994), semi-synthesis from its precursor (Commercn et al., 1995), plant tissue or cell culture (Hu et al., 2003). Endophytic fungus from various taxus species produces taxol although the amounts are very small. First taxol producing endophytic fungi T. andreanae is from Taxus brevifolia (Stierle et al., 1993). Since then, many other endophytic fungi from other plant species have been successfully reported to produce taxol (Strobel et al., 1996; Li et al., 1996). Tylophora indica (Burm. f) (Asclepiadaceae) is a climbing perennial plant that grows in India, commonly called as antmool in Ayurveda. The leaves of Tylophora have been traditionally used as a folk medicine. It has been used for respiratory problems such as asthma, allergies, bronchitis and common cold. The roots and leaves contain 0.2 to 0.46% of therapeutically important alkaloids namely tylophorine, tylophorinine and tylophrinidine. Major alkaloid tylophorine has immunosuppressive, anti-inflammatory (Gopalakrishnan et al., 1979), anti-tumor (Donaldson et al., 1968) and anti-amoebic (Bhutani et al., 1987) properties. Since, T. indica is an endemic plant an attempt is made to conserve the medicinal plant through exploration of endophytic fungi and screened for its taxol production. Materials and methods Plant collection: Tylophora indica fresh plants were collected from Kerala Forest Research Institute (KFRI), Trissur, Kerala, South India during the month of January 2012 (Fig. 1). The taxonomical identity of the plant was confirmed by Dr. D. Narashiman, Department of Botany, Madras Christian College, Tambaram, South India. Fig 1. Tylophora indica. RESEARCH ARTICLE