In vitro propagation of brown oak (Quercus semecarpifolia Sm.) from seedling explants Sushma Tamta & Lok Man S. Palni & Vijay K. Purohit & Shyamal K. Nandi Received: 3 September 2007 / Accepted: 21 March 2008 / Published online: 10 June 2008 / Editor: Florent Engelmann # The Society for In Vitro Biology 2008 Abstract An efficient and reproducible method for the regeneration of multiple shoots of brown oak (Quercus semecarpifolia Sm.) has been developed in which a part of the petiolar tube containing a primary shoot is used as the explant. Explants derived from in vitro grown seedlings were cultured either on Murashige and Skoog or Woody Plant medium (WPM) containing different concentrations of benzyladenine (BAP) throughout the range of 1–20 μM. WPM supplemented with 20 μM BAP was found to be best for adventitious shoot induction and for the multiplication of individual shoots. In-vitro-produced shoots were rooted using a two-step method. Firstly, shoots were cultured on WPM containing indolebutyric acid (IBA) at either 50 or 100 μM for 24 or 48 h. Secondly, the shoots were transferred to plant-growth-regulator-free half-strength WPM. The second step not only considerably improved the rooting percentage but also minimized the formation of basal callus. The most effective first-step treatment was found to be 100 μM IBA for 24 h, which initiated rooting at a frequency of 100%. Well-rooted plants were transferred to plastic cups containing nonsterile, sieved soil and farmyard manure, hardened under greenhouse conditions, and then successfully established in pots. This procedure is suitable for use in large-scale production of plants and may have potential application in additional oak species. Keywords Brown oak . Petiolar tube . Quercus semecarpifolia . Rooted plants . Shoot regeneration Introduction Quercus semecarpifolia Sm. (family Fagaceae), commonly known as brown or kharsu oak, predominates as a forest- forming species at an elevation of around 2,400 m in the Indian Himalayas (Singh and Singh 1987). It is a source of firewood, charcoal, fodder, and agricultural implements and is used in tasar (silk) sericulture (Singh et al. 1997). Additionally, it plays an important role in water conserva- tion and in the prevention of soil erosion. A result of these multiple uses has been the indiscriminate harvesting of fully grown trees and a significant decline in population numbers of Q. semecarpifolia. Natural regeneration through seed is known to be poor (Troup 1921). Vegetative propagation through rooted stem cuttings has, in general, not been successful in oaks (Chalupa 1982; Bhardwaj et al. 1996; Tamta et al. 2000) and thus far has not been reported for Q. semecarpifolia. Tissue culture and micropropagation offer a potential means for rapid multiplication of forest trees as well as for the conservation of rare species and has been successfully applied in a number of Himalayan tree species (Bisht et al. 1998). In vitro propagation via axillary shoot multiplication In Vitro Cell.Dev.Biol.-Plant (2008) 44:136–141 DOI 10.1007/s11627-008-9138-x S. Tamta : L. M. S. Palni : V. K. Purohit : S. K. Nandi G. B. Pant Institute of Himalayan Environment and Development, Kosi-Katarmal, Almora 263 643, U. A., India S. Tamta Botany Department, D.S.B. Campus, Kumaun University, Nainital, Uttarakhand 263 002, India L. M. S. Palni State Biotechnology Programme, Government of Uttaranchal, Biotec Bhavan, P.O. Haldi, Pantnagar 263 146 Uttarakhand, India S. Tamta (*) Kumaun University, Nainital, Uttarakhand 263 002, India e-mail: sushma_tamta@yahoo.com