The isopentenyl transferase gene is effective as a selectable marker gene for plant transformation in tobacco (Nicotiana tabacum cv. Petite Havana SRI)

• Published in: Plant cell reports 2001-01, Vol.20 (1), p.60-66
• Main Authors: ENDO, S, KASAHARA, T, SUGITA, K, MATSUNAGA, E, EBINUMA, H
• Format: Article
• Language: English
• Subjects: Antibiotics; Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; Genes; Genetic engineering; Genetic technics; isopentenyl transferase; kanamycin; Methods. Procedures. Technologies; Nicotiana tabacum; Transgenic plants; Vectors (cloning, transfer, expression). Insertion sequences and transposons
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/s002990000279

A selection method for transformed cells which does not inhibit regeneration is important for the establishment and optimization of a transformation protocol. We have assessed the 35S-ipt gene from Agrobacterium tumefaciens as a selectable marker gene. The identification of ipt-expressing cells from nontransformed cells enabled morphological selection without the use of kanamycin and also allowed for the elimination of a high proportion of nonexpressing cells. Ipt selection of tobacco leaf discs (Nicotiana tabacum cv. Petite Havana SRI) resulted in a 2.7-fold higher transformation frequency compared to kanamycin selection. Overexpression of the ipt gene favored plant regeneration from transformed cells, and the transformation frequency of the ipt plus kanamycin selection resulted in a 1.6-fold higher transformation frequency than kanamycin selection alone. These results indicate that this procedure might provide a strategy whereby transgenic plants can be efficiently obtained and some of the problems related to the use of antibiotics diminished.