caa a22 4500 467900728 CHVBK 20180406152817.0 cr unu---uuuuu 170328e20060301xx s 000 0 eng 10.1007/s00299-005-0045-x doi (NATIONALLICENCE)springer-10.1007/s00299-005-0045-x Stable transformation and direct regeneration in Coffea canephora P ex. Fr. by Agrobacterium rhizogenes mediated transformation without hairy-root phenotype [Elektronische Daten] [Vinod Kumar, K. Satyanarayana, S. Sarala Itty, E. Indu, P. Giridhar, A. Chandrashekar, G. Ravishankar] A system for genetic transformation of Coffea canephora by co-cultivation with Agrobacterium rhizogenes harbouring a binary vector has been developed. The objective of the present study was the genetic transformation and direct regeneration of transformants through secondary embryos bypassing an intervening hairy root stage. Transformants were obtained with a transformation efficiency up to 3% depending on the medium adjuvant used. A. rhizogenes strain A4 harbouring plasmid pCAMBIA 1301 with an intron uidA reporter and hygromycin phosphotransferase (hptII) marker gene was used for sonication-assisted transformation of Coffea canephora. The use of hygromycin in the secondary embryo induction medium allowed the selection of transgenic secondary embryos having Ri T-DNA along with the T-DNA from the pCAMBIA 1301 binary vector. In addition transgenic secondary embryos devoid of Ri-T-DNA but with stable integration of the T-DNA from the binary vector were obtained. The putative transformants were positive for the expression of the uidA gene. PCR and Southern blot analysis confirmed the independent, transgenic nature of the analysed plants and indicated single and multiple locus integrations. The study clearly demonstrates that A. rhizogenes can be used for delivering transgenes into tree species like Coffea using binary vectors with Agrobacterium tumefaciens T-DNA borders. Springer-Verlag, 2005 Coffea canephora nationallicence Agrobacterium rhizogenes nationallicence Genetic transformation nationallicence Secondary somatic embryogenesis nationallicence Kumar Vinod Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut Satyanarayana K. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut Sarala Itty S. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut Indu E. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut Giridhar P. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut Chandrashekar A. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut Ravishankar G. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut Plant Cell Reports Springer-Verlag 25/3(2006-03-01), 214-222 0721-7714 25:3<214 2006 25 299 https://doi.org/10.1007/s00299-005-0045-x text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-005-0045-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kumar Vinod Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut NATIONALLICENCE 700 1- Satyanarayana K. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut NATIONALLICENCE 700 1- Sarala Itty S. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut NATIONALLICENCE 700 1- Indu E. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut NATIONALLICENCE 700 1- Giridhar P. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut NATIONALLICENCE 700 1- Chandrashekar A. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut NATIONALLICENCE 700 1- Ravishankar G. Plant Cell Biotechnology Department, Central Food Technological Research Institute, 570 020, Mysore, Karnataka, India aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/3(2006-03-01), 214-222 0721-7714 25:3<214 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer