Phyton (Buenos Aires)
versión On-line ISSN 1851-5657
SANCHEZ, N; MANZANERA, JA y BUENO, MA. Agrobacterium-mediated transformation of white poplar (Populus alba L.): (with 3 figures & 1 table). Phyton (B. Aires) [online]. 2004, vol.73, pp. 123-130. ISSN 1851-5657.
A simple method is presented for the genetic transformation of white poplar (Populus alba L.). Root suckers and buds from branches of adult trees were established in vitro. Callus was induced on injured leaves cultured on medium supplemented with 0.45 µM thidiazuron and 0.1 µM a-naphthalene acetic acid. Non-transformed leaves and calli did not survive concentrations higher than 85 µM kanamycin or 18 mM hygromycin. The leaf explants were inoculated with the disarmed Agrobacterium tumefaciens LBA4404/p35S GUS INT/pCAMBIA 2301 strain. Twenty % of the cultured callus explants was kanamycin-resistant two months after the co-inoculation experiment. The rate of adventitious bud induction on resistant calli was 17 %. The analysis of b-glucuronidase expression two months after the co-cultivation experiment and the DNA amplification of a 2500 bp band with 35S sense primer and NOS antisense primer confirmed the transformation of 100 % kanamycin-resistant calli and 4 % adventitious shoots. This method forms a basis for genetic transformation of white poplar.
Palabras llave : BAP; benzil aminopurine; TDZ, Thidiazuron; IBA, indole butyric acid; NAA, naphthalene acetic acid; KAN, kanamycin.