Transformation and regeneration of Nicotiana edwardsonii

• Published in: Plant science (Limerick) Vol. 64; no. 1; pp. 67 - 78
• Main Authors: Kiernan, J.M., Goldberg, K.-B., Young, M.J., Schoelz, J.E., Shepherd, R.J.
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier Ireland Ltd 1989; Elsevier Science
• Subjects: Agrobacterium tumefaciens; Agronomy. Soil science and plant productions; Biological and medical sciences; caulimoviruses; Classical genetics, quantitative genetics, hybrids; Economic plant physiology; Fundamental and applied biological sciences. Psychology; Genetics of eukaryotes. Biological and molecular evolution; In vitro culture; Nicotiana clevelandii; Nicotiana edwardsonii; Nicotiana glutinosa; Pteridophyta, spermatophyta; tissue culture; Vegetals; tissue culture; MS; NPT II; Agrobacterium tumefaciens; IAA; bp; FMV; NAA; caulimoviruses; BAP; Nicotiana clevelandii; Nicotiana edwardsonii; CaMV; 2iP; Nicotiana glutinosa; Culture medium; Nicotiana; Genetic transformation; Genetic marker; Tissue culture; Regeneration; Antibiotic; Sensitivity resistance; Dicotyledones; Angiospermae; Immunoblotting assay; Bacteria; Spermatophyta; Transgenic plant; Rhizobiaceae; Solanaceae; Southern blotting; Shoot
• ISSN: 0168-9452; 1873-2259
• DOI: 10.1016/0168-9452(89)90153-2

Several previously developed media for regeneration of Nicotiana species were evaluated for their ability to regenerate shoots of Nicotiana edwardsonii, a fertile amphidiploid of N. glutinosa × N. clevelandii. Two media were superior to the others, producing shoots within 3–4 weeks. Roots were easily regenerated on a simple rooting medium. Ti plasmid- Agrobacterium tumefaciens mediated DNA transfer was used to genetically transform N. edwardsonii with gene VI of strains D4 and CM1841 of cauliflower mosaic virus (CaMV) and strain Sc3 of figwort mosaic virus (FMV). When using gene vectors with the neomycin phosphotransferase type II gene and antibiotics kanamycin or G418, a related antibiotic, in a positive selection procedure, a marked inhibition of shoot formation was observed. Consequently, it was necessary to use much lower concentrations of the antibiotics in the selection media to obtain transgenic plants than are used for other species such as N. tabacum and Datura innoxia. Insertion of the desired caulimoviral genes was demonstrated by the presence of the appropriate DNA sequences and the corresponding protein products. The level of expression of the inserted genes is briefly compared to D. innoxia transformed with the same genes. The authors believe this to be a first report of the transformation and regeneration of N. edwardsonii, a widely used host of plant viruses.