Pakistan Journal of Biological Sciences1028-88801812-5735Asian Network for Scientific Information10.3923/pjbs.2014.472.481hptII)]]>HtweNwe Nwe LingHo Chai ZamanFaridah Qamaruz MaziahMahmood 42014174Rice is one of the most important cereal crops with great potential for biotechnology progress. In transformation method, antibiotic resistance genes are routinely used as powerful markers for selecting transformed cells from surrounding non-transformed cells. In this study, the toxicity level of hygromycin was optimized for two selected mutant rice lines, MR219 line 4 and line 9. The mature embryos were isolated and cultured on an MS medium with different hygromycin concentrations (0, 20, 40, 60, 80 and 100 mg L-1). Evidently, above 60 mg L-1 was effective for callus formation and observed completely dead. Further there were tested for specific concentration (0-60). Although, 21.28% calli survived on the medium containing 45 mg L-1 hygromycin, it seemed suitable for the identification of putative transformants. These findings indicated that a system for rice transformation in a relatively high frequency and the transgenes are stably expressed in the transgenic plants. Green shoots were regenerated from the explant under hygromycin stress. RT-PCR using hptII and gus sequence specific primer and Southern blot analysis were used to confirm the presence of the transgene and to determine the transformation efficiency for their stable integration in regenerated plants. This study demonstrated that the hygromycin resistance can be used as an effective marker for rice transformation.]]>Akutsu, M., T. Ishizaki and H. Sato,2004Agrobacterium tumefaciens.]]>22561568Aoki, T., A. Kamizawa and S. Ayabe,2002Agrobacterium-mediated transformation of Lotus japonicus with reliable antibiotic selection.]]>21238243Brodersen, D.E., W.M. Jr, Clemons, A.P. Carter, R.J. Morgan-Warren, B.T. Wimberly and V. Ramakrishnan,200010311431154Cho, M.J., W. Jiang and P.G. Lemaux,1999148917Datta, S.K., A. Peterhans, K. Datta and I. Potrykus,1990Indica-rice plants recovered from protoplasts.]]>8736740Datta, K., A. Vasquez, J. Tu, L. Torrizo and M.F. Alam et al.,1998972030Eustice, D.C. and J.M. Wilhelm,1984265360Gritz, L. and J. Davies,1983Escherichia coli and Saccharomyces cerevisiae.]]>25179188Hille, J., F. Verheggen, P. Roelvink, H. Franssen, A. Van Kammen and P. Zabel,19867171176Janna, O.A., M. Maziah and G.K.A. Parvezz,200088593Liau, C.H., S.J. You, V. Prasad, H.H. Hsiao, J.C. Lu, N.S. Yang and M.T. Chan,2003Agrobacterium tumefaciens-mediated transformation of an Oncidium orchid.]]>21993998Liu, Q.C., H. Zhai, Y. Wang and D.P. Zhang,200137564567McBride, K.E. and K.R. Summerfelt,199014269276Men, S., X. Ming, Y. Wang, R. Liu, C. Wei and Y. Li,200321592598Miki, B. and S. McHugh,2004107193232Mishiba, K.I., K. Ishikawai, O. Tsujii and M. MII,2001Lavandula latifolia medicus) mediated by Agrobacterium.]]>75287292Mulsant, P., A. Gatignol, M. Dalens and G. Tiraby,198814243252Nakazawa, M. and M. Matsui,2003342830Parveez, G.K.A., M.K.U. Chowdhury and N.M. Saleh,1996Elaeis guineensis Jacq. transformation.]]>4219228Rao, S.S., L. Mamadou, M. McConnell, R. Polisetty, P. Kwanyuen and D. Hildebrand,20092009Roa-Rodriguez, C., C. Nottenburg and D. Aston,20032003Schroeder, H.E., A.H. Schotz, T. Wardley-Richardson, D. Spencer and T.J.V. Higgins,1993Pisum sativum L.).]]>101751757Shaw, J.F., H.H. Chen, M.F. Tsai, C.I. Kuo and L.C. Huang,2002Brassica oleracea.]]>9211261Suzuki, S., K. Supaibulwatana, M. Mii and M. Nakano,2001Agapanthus praecox ssp. orientalis (Leighton) Leighton via Agrobacterium-mediated transformation of embryogenic calli.]]>1618997Tee, C.S., M. Marziah, C.S. Tan and M.P. Abdullah,20012001pp: 687690Tian, L., F.A. Canli, X. Wang and S. Sibbald,2009Prunus domestica L. using the hpt gene coding for hygromycin resistance as the selectable marker. L.]]>119339343Tian, L., M. Jordan and B. Miki,20062006pp: 9-20pp: 9-20Yu, Z., M. Chen, L. Nie, H. Lu and X. Ming et al.,1999588792Zheng, Z., A. Hayashimoto, L. Zhijian and N. Murai,199197832835Jefferson, R.A.,1989gus reporter gene system.]]>342837838