Cucumber (Cucumis sativus L.).
We describe two novel Agrobacterium tumefaciens-based methods of cucumber transformation. The first involves direct regeneration from leaf microexplants selected on kanamycin-containing medium. The second involves regeneration from a long-term established embryogenic suspension culture emitting green autofluorescence (GAF) and selection on medium containing hygromycin. In the latter method, GAF was used as a reporter, thereby allowing a simple and reliable identification of transgenic cells with a high regeneration capacity. (No false positives were observed.) The transformation efficiency in the leaf microexplants fluctuated from 0.8 to 6.5% of the primary explants, whereas in the embryogenic suspension-cultured cells it varied from 6.4 to 17.9% of the aggregates. In the GAF method, the step involving the elimination of the Agrobacterium cells by antibiotics could be omitted; however, this reduced the transformation efficiency to about 3%. The time required from inoculation to regenerated plant in the greenhouse was the same for both methods, but the GAF method required more preinoculation time than the leaf microexplant method.
Burza
Wojciech
W
Warsaw Agricultural University, Department of Plant Genetics Breeding and Biotechnology, Warsaw, Poland.
Zuzga
Sabina
S
Yin
Zhimin
Z
Malepszy
Stefan
S
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D018553:Cucumis sativus / Q000166:cytology / Q000196:embryology / Q000235:genetics* / Q000382:microbiology; D004351:Drug Resistance / Q000235:genetics; D018014:Gene Transfer Techniques; D005819:Genetic Markers; D018515:Plant Leaves / Q000166:cytology / Q000196:embryology / Q000235:genetics* / Q000382:microbiology; D030821:Plants, Genetically Modified / Q000166:cytology / Q000196:embryology / Q000235:genetics* / Q000382:microbiology; D014170:Transformation, Genetic
D005819:Genetic Markers