caa a22 4500 463235754 CHVBK 20180405153256.0 cr unu---uuuuu 170326e20070301xx s 000 0 eng 10.1007/s11240-006-9199-7 doi (NATIONALLICENCE)springer-10.1007/s11240-006-9199-7 Pinto-Sintra A. Department of Genetics & Biotechnology, Centre of Genetics and Biotechnology (CGB), University of Trás-os-Montes and Alto Douro, P.O. Box 1013, 5001-801, Vila Real, Portugal aut Establishment of embryogenic cultures and plant regeneration in the Portuguese cultivar ‘Touriga Nacional' of Vitis vinifera L [Elektronische Daten] [A. Pinto-Sintra] ‘Touriga Nacional' is the most important Portuguese grapevine cultivar used for Port wine, table wine and varietal wine production. In order to obtain a reproducible plant regeneration system that allows the application of biotechnological tools to grapevine breeding, embryogenic cultures were induced from immature flowers of three Touriga Nacional selected clones. Gynoecia and anthers were cultured on Nitsch and Nitsch (Science 163:85-87, 1969) basal medium supplemented with four combinations of the growth regulators 6-benzylaminopurine (BAP), 2,4-dichlorophenoxyacetic acid (2,4-D) and indole-3-acetyl-l-aspartic acid (IASP), at 28°C, in the dark. Primary callus, observed on anthers and gynoecia in all media, produced embryogenic callus when cultured on differentiation medium, at 24°C under light. The efficiency on induction of embryogenic callus ranged from 1.2±4.7% to 7.9±13.8% in anthers, and from 17.9±24.9% to 25.3±22.9% in gynoecia. Seven lines of embryogenic cultures were established from the three clones. Multiplication of embryogenic calluses was successfully obtained in maintenance medium, at 26°C, in the dark. These embryogenic calluses produced somatic embryos when subcultured on differentiation medium, under a 16h photoperiod. Somatic embryos were isolated and cultured on germination medium to achieve conversion which ranged from 35.3±48.5% to 72.7±45.6%. The plantlets obtained were cultured in medium without growth regulators. Secondary embryogenesis was also frequently observed in the hypocotyl-root transition region of somatic embryos. Although some morphological variation occurred between somatic embryos, the regenerated plantlets had a normal phenotype. Maintenance of embryogenic cultures has been achieved since 2002. Springer Science+Business Media B.V., 2007 Grapevine nationallicence Anthers nationallicence Gynoecia nationallicence Embryogenic calluses nationallicence Somatic embryos nationallicence Secondary embryogenesis nationallicence SEM nationallicence 2,4-D : 2,4-Dichlorophenoxyacetic acid nationallicence BAP : 6-Benzylaminopurine nationallicence CIM : Callus initiation medium nationallicence FAO : Food and Agriculture Organization nationallicence Ha : Hectare nationallicence IASP : Indole-3-acetyl-l-aspartic acid nationallicence MS : Murashige and Skoog (1962) nationallicence Mt : Metric ton nationallicence NN : Nitsch and Nitsch (1969) nationallicence SEM : Scanning electron microscope nationallicence TN : ‘Touriga Nacional' nationallicence Plant Cell, Tissue and Organ Culture Kluwer Academic Publishers 88/3(2007-03-01), 253-265 0167-6857 88:3<253 2007 88 11240 https://doi.org/10.1007/s11240-006-9199-7 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11240-006-9199-7 text/html Onlinezugriff via DOI NATIONALLICENCE 100 1- Pinto-Sintra A. Department of Genetics & Biotechnology, Centre of Genetics and Biotechnology (CGB), University of Trás-os-Montes and Alto Douro, P.O. Box 1013, 5001-801, Vila Real, Portugal aut NATIONALLICENCE 773 0- Plant Cell, Tissue and Organ Culture Kluwer Academic Publishers 88/3(2007-03-01), 253-265 0167-6857 88:3<253 2007 88 11240 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer