American Journal of Plant Physiology1557-45391557-4547Academic Journals Inc.10.3923/ajpp.2007.333.343Pinus roxburghii]]>MalabadiRavindra B. NatarajaK. 6200726Present research highlights for the first time the expression of cDNA clones of genes involved in programming the apical meristem cells towards somatic embryogenic pathway influenced by external environmental stimulus like cold-pretreatment. Differential display was used to isolate the genes which are expressed specifically in embryogenic tissue induced by cold-pretreatment of thin sections of vegetative shoot apices of mature trees of Pinus roxburghii. We have developed a rapid method which employs magnetic beads to capture mRNA from cell lysates. Of the 56 cold-enhanced embryogenic-associated cDNAs identified, 20 were cloned. Nine of the 20 fragments which generated single bands on re-amplification were selected for cloning and further analysis. During reverse northern hybridization, all the 20 clones selected generated a positive signal when probed with labeled cDNA from cold-enhanced embryogenic tissue, but no signal when probed with cDNA from the non-embryogenic tissue (control treatment). All the 20 clones thus contained inserts that were specific to cold-enhanced somatic embryogenesis. This approach allows us to perform differential display and construction of cDNA libraries from the small amount of embryogenic tissue and outline a PCR-based method for confirming differential expression of large number of cloned bands in cases where RNA quantities are limiting.]]>Bishop-Hurley, S.L., R.C. Gardner and C. Walter,2003Pinus radiata.]]>74267281Dong, J.Z. and D.I. Dunstan,1996Picea glauca).]]>2008591Dong, J.Z. and D.I. Dunstan,1996Picea glauca).]]>199459466Dong, J.Z. and D.I. Dunstan,1997Picea glauca.]]>201189194Dong, J.Z. and D.I. Dunstan,1999Picea glauca and their comparative expression during zygotic embryogenesis.]]>39859864Feliciello, I. and G. Chinali,1993Escherichia coli.]]>212394401Goldberg, R.B., G. De-Paiva and R. Yadegari,1994266605614Goncalves, S., J. Cairney, M. Oliveira and C. Miguel,2005Pinus pinaster).]]>13203216Goncalves, S., J. Cairney, J. Maroco, M. Oliveira and C. Miguel,2005222556563Gupta, P.K. and D.J. Durzan,19854177179Lefebvre, P.A., C.D. Silflow, E.D. Wieben and J.L. Rosenbaum,1980Chlamydomonas flagella.]]>20467477Liang, P. and A.B. Pardee,1992257967971Lorenz, W.W., F. Sun, C. Liang, D. Kolychev and H. Wang et al.,2005Pinus taeda) roots identified by analyses of expressed sequence tag libraries.]]>26116Malabadi, R.B. and J. Van Staden,2003Pinus patula trees.]]>69450451Malabadi, R.B., H. Choudhury and P. Tandon,2004Pinus kesiya (Royle ex. Gord) promoted by partial desiccation and Gellan gum.]]>102449459Malabadi, R.B. and J. Van Staden,2005Pinus patula.]]>251116Malabadi, R.B. and J. Van-Staden,2005Pinus patula.]]>41181186Malabadi, R.B. and J. Van-Staden,2005Pinus patula trees.]]>82259265Malabadi, R.B. and J. Van Staden,20052005Malabadi, R.B.,2006Pinus roxburghii.]]>193538Malabadi, R.B. and J. Staden-Van,2006Pinus patula is mediated by calcium.]]>72613618Malabadi, R.B. and K. Nataraja,2006Pinus roxburghii Sarg. Trees.]]>42152159Malabadi, R.B. and K. Nataraja,2006Pinus roxburghii SARG.]]>6114120Malabadi, R.B. and K. Nataraja,2007Pinus wallichiana A.]]>24553Malabadi, R.B. and K. Nataraja,2007Pinus roxburghii sarg.]]>34047Maniatis, T., E.F. Fritsch and J. Sambrock,19823rd Edn.,Mathieu, M., M.A. Lelu-Walter, A.S. Blervacq, H. David, S. Hawkins and G. Neutelings,200661615627Mou, L., H. Miller, J. Li, E. Wang and L. Chalifour,1994199564569Pullman, G.S., S. Johnson, G. Peter, J. Cairney and N. Xu,200321747758Salzman, R.N., T. Fujita, K. Zhu-Salzman, P.M. Hasegawa and R.A. Bressan,1999171117Stasolla, C., L.V. Zyl, U. Egertsdotter, D. Craig, W. Liu and R.R. Sederoff,20031314960Wan, C.Y. and T.A. Wilkins,1994Gossypium hirsutum L.).]]>223712Xu, N., B. Johns, G. Pullman and J. Cairney,199715377391