Journal of Medicinal Plants Research Vol. 5(28), pp. 6460-6467, 30 November, 2011 Available online at http://www.academicjournals.org/JMPR ISSN 1996-0875 ©2011 Academic Journals DOI: 10.5897/JMPR11.146 Full Length Research Paper An efficient in vitro regeneration protocol for faba bean (Vicia faba L.) Firoz Anwar 1 *, Salem S. Alghamdi 1 , Megahed H. Ammar 1 and K. H. M. Siddique 1,2 1 Legume Research Unit, Plant Production Department, Faculty of Food and Agricultural Sciences, King Saud University, P. O. Box 2460, Riyadh 11451, Saudi Arabia. 2 The UWA Institute of Agriculture, The University of Western Australia, 35 Stirling Highway, Crawley WA 6009, Australia. Accepted 11 April, 2011 A reliable regeneration system for faba bean (Vicia faba L.) has been difficult to establish, delaying its genetic improvement. In the present work a rapid, reproducible and efficient regeneration method was developed for faba bean using single cotyledon explants with half embryonal axis. MS medium supplemented with 6 μM TDZ (thidiazuron), 10 μM 2-iP and 4 μM kinetin induced 30 to 50 adventitious buds /shoots after two weeks of culture, which were elongated on MS medium supplemented with 6 μM 2-iP and 2 μM kinetin. With healthy and strong roots established, shoots were transferred to MS medium supplemented with 5 μM IBA within 10 to 14 days. Shoots of 5 cm long were most suitable for rooting. Potting-mixture with good aeration and less capacity to retain water was most suitable for successful establishment of faba bean plantlets. Garden soil mixed with sand (gravel) and bio-manure (1:1:1) was most suitable for transplantation. TDZ promoted adventitious bud formation while 5 μM IBA was most suitable for rooting, higher concentrations were toxic to plantlets. Aeration of the potting mixture was important for rapid micropropagation and successful establishment. The efficient regeneration protocol reported here allows for successful micropropagation of faba bean, which is essential for future genetic improvement of plants via transformation protocols. Key words: Vicia fab, organogenesis, transplantation, TDZ, IBA and kinetin. INTRODUCTION Legumes (Leguminosae) are the third largest family of dicotyledons. Faba bean (Vicia faba L.) belongs to the Fabaceae family and has many common names. It is one of the most important grain legumes in the world since it is an excellent source of protein, used for both human consumption and animal feed. Faba bean also plays an important role in biological fixation of aerial nitrogen (Duke 1981; Jelenić et al., 2000). This specie alone occupied nearly 3.2 x 10 6 ha worldwide in 1991 (FAO statistics, 1992) with a world production close to 4.5 million tons in 2004 (Gutierrez, 2006). Unfortunately, faba bean is susceptible to environmental conditions, biotic and abiotic stresses, and *Corresponding author. E-mail: anfiroz@gmail.com. has unstable yields. Difficulties in pollination control and a limited genetic pool has led to slow progress in crop varietal improvement (Bond, 1987; Bond et al., 1985; Selva et al., 1989). Legumes such as faba bean appear to be recalcitrant in in vitro regeneration (Khalafalla and Hattori, 2000; Anwar, 2007; Anwar et al., 2008, 2010), due to difficulties regenerating from callus and the release of phenolic compounds. As a result, studies on in vitro culture of faba bean are difficult (Böttinger et al., 2001). Regeneration of faba bean via indirect somatic embryogenesis was reported by Griga et al. (1987) using cotyledons on media supplemented with 2,4-D. Somatic embryos at the globular to early torpedo stage were released, but their development terminated at the late torpedo stage. In tissue culture, regenerated plants from cultured cells are known to exhibit genetic and epigenetic changes collectively called somaclonal variations