ISSN:
1573-5044
Keywords:
Agrobacterium genetic transformation
;
inoculation temperature
;
Malux × domestica
;
plasmids
;
rooting
;
stem elongation
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract Transformed shoots of the major apple scion cultivar ‘Delicious’ (Malus × domestica Borkh.) were obtained by cocultivation withAgrobacterium tumefaciens carrying disarmed plasmids. The transformation efficiency was influenced by the type of plasmid and by the inoculation temperature. Initial selection involved a callus stage followed by shoot regeneration. Shoot regeneration occurred only in the dark. Shoots grew in the light and were rooted in the presence of 100 mg l−1 kanamycin. Of the range of plasmids tested, the cointegrates pGV 3850::1103neo and pGV 3850::1103gus gave a higher frequency of transformation than the binary vector pGV 3111 × pKIWI. Elongation of transformed shoots was enhanced by culture in a mixture of the cytokinins 6(γ-γ-dimethylallylamino)purine and 6-benzyladenine. Up to 60% of the elongated shoots rooted in 100 mg l−1 kanamycin. Transformation was indicated by kanamycin resistance, β-glucuronidase assay, nopaline synthesis, and by integration of the T-DNA as judged by Southern analysis.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF00046089
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