caa a22 4500 463236599 CHVBK 20180405153259.0 cr unu---uuuuu 170326e20071001xx s 000 0 eng 10.1007/s11240-007-9278-4 doi (NATIONALLICENCE)springer-10.1007/s11240-007-9278-4 Evaluation of MAT-vector system in white poplar ( Populus alba L.) and production of ipt marker-free transgenic plants by ‘single-step transformation' [Elektronische Daten] [Samanta Zelasco, Valentina Ressegotti, Massimo Confalonieri, Daniela Carbonera, Paolo Calligari, Martina Bonadei, Stefano Bisoffi, Keiko Yamada, Alma Balestrazzi] Genetic transformation of an elite white poplar genotype (Populus alba L., cv. ‘Villafranca') was performed with MAT vectors carrying the ipt and rol genes from Agrobacterium spp. as morphological markers. The effects associated with the use of different gene promoters and distinct invitro regeneration protocols were evaluated. Poplar plantlets showing abnormal ipt and rol phenotypes were produced only in the presence of exogenous growth regulators. The occurrence of abnormal ipt and rol phenotypes allowed the visual selection of transformants. The ipt-type MAT vector pEXM2 was used to monitor the activity of the yeast site-specific recombination R/RS system in the transformed white poplar cells. Results from these experiments demonstrated that recombinase-mediated excision events occurred during the early stages of invitro culture, thus causing the direct production of ipt marker-free transgenic plants with normal phenotype at an estimated frequency of 36.4%. Beside this unexpected finding, transgenic ipt-shooty plants were obtained at a frequency of 63.6% and normal shoots were subsequently recovered after a prolonged period of invitro culture. Although the transformation efficiency observed in this study, using both ipt and nptII genes as selection markers, was similar to that previously reported with standard vectors carrying only the nptII gene, the easy identification of ipt transformants, the early recombinase-mediated excision events and finally the relatively short time period required to produce ipt marker-free transgenic plants support for the choice of MAT vectors as a reliable strategy for the future production of marker-free GM poplars. Springer Science+Business Media B.V., 2007 ipt nationallicence Marker-free transgenic plant nationallicence Morphological marker nationallicence Populus alba L nationallicence Recombinase nationallicence rol nationallicence Single-step transformation nationallicence BAP : Benzyl-aminopurine nationallicence NAA : α-Naphthaleneacetic acid nationallicence TDZ : Thidiazuron nationallicence Zelasco Samanta Istituto di Sperimentazione per la Pioppicoltura—C.R.A, via di Frassineto 35, 15033, Casale Monferrato, AL, Italy aut Ressegotti Valentina Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100, Pavia, Italy aut Confalonieri Massimo Istituto Sperimentale per le Colture Foraggere—C.R.A, viale Piacenza 29, 26900, Lodi, Italy aut Carbonera Daniela Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100, Pavia, Italy aut Calligari Paolo Istituto di Sperimentazione per la Pioppicoltura—C.R.A, via di Frassineto 35, 15033, Casale Monferrato, AL, Italy aut Bonadei Martina Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100, Pavia, Italy aut Bisoffi Stefano Istituto di Sperimentazione per la Pioppicoltura—C.R.A, via di Frassineto 35, 15033, Casale Monferrato, AL, Italy aut Yamada Keiko Nippon Paper Industries, CO., LTD, 5-21-1, Oji, Kita-Ku, 114-0002, Tokyo, Japan aut Balestrazzi Alma Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100, Pavia, Italy aut Plant Cell, Tissue and Organ Culture Springer Netherlands 91/1(2007-10-01), 61-72 0167-6857 91:1<61 2007 91 11240 https://doi.org/10.1007/s11240-007-9278-4 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11240-007-9278-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zelasco Samanta Istituto di Sperimentazione per la Pioppicoltura—C.R.A, via di Frassineto 35, 15033, Casale Monferrato, AL, Italy aut NATIONALLICENCE 700 1- Ressegotti Valentina Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100, Pavia, Italy aut NATIONALLICENCE 700 1- Confalonieri Massimo Istituto Sperimentale per le Colture Foraggere—C.R.A, viale Piacenza 29, 26900, Lodi, Italy aut NATIONALLICENCE 700 1- Carbonera Daniela Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100, Pavia, Italy aut NATIONALLICENCE 700 1- Calligari Paolo Istituto di Sperimentazione per la Pioppicoltura—C.R.A, via di Frassineto 35, 15033, Casale Monferrato, AL, Italy aut NATIONALLICENCE 700 1- Bonadei Martina Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100, Pavia, Italy aut NATIONALLICENCE 700 1- Bisoffi Stefano Istituto di Sperimentazione per la Pioppicoltura—C.R.A, via di Frassineto 35, 15033, Casale Monferrato, AL, Italy aut NATIONALLICENCE 700 1- Yamada Keiko Nippon Paper Industries, CO., LTD, 5-21-1, Oji, Kita-Ku, 114-0002, Tokyo, Japan aut NATIONALLICENCE 700 1- Balestrazzi Alma Dipartimento di Genetica e Microbiologia, Università di Pavia, via Ferrata 1, 27100, Pavia, Italy aut NATIONALLICENCE 773 0- Plant Cell, Tissue and Organ Culture Springer Netherlands 91/1(2007-10-01), 61-72 0167-6857 91:1<61 2007 91 11240 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer