ISSN:
1573-5044
Keywords:
antibiotics
;
cocultivation
;
organogenesis
;
Populus
;
tissue culture
;
thidiazuron
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract A method for Agrobacterium-mediated transformation of hybrid poplar (Populus alba x P. grandidentata cv. ‘Crandon’) suspension cultures and regeneration of transformed plants is described. Transformants were recovered when suspension cultures were inoculated with Agrobacterium tumefaciens at a density of 107 colony-forming units ml-1, cocultivated for 48 h, and plated to cellulose acetate filters on Woody Plant Medium containing 4.5 μM 2,4-dichlorophenoxyacetic acid and 250 mg l-1 cefotaxime. Levels of cefotaxime greater than 250 mg l-1 were unnecessary for control of residual bacteria and inhibited callus growth. Transgenic plants were regenerated by culturing the transformed callus on media containing 0.11 to 27 μM thidiazuron. In contrast to thidiazuron, N6-benzyladenine had a negative effect on shoot regeneration; the callus became necrotic when we attempted to induce shoots with concentrations of 1.1 to 8.9 μM, and growth was inhibited when concentrations of 0.11 or 0.22 μM were used to regenerate callus from suspension cultures. Following cocultivation of poplar suspension cultures, we recovered transgenic plants containing the maize transposon Ac, and callus containing an insect toxin gene from Bacillus thuringiensis.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00048316
Permalink
