GUS expression in blueberry (Vaccinium spp.): factors influencing Agrobacterium-mediated gene transfer efficiency
1998
Cao, X. | Liu, Q. | Rowland, L.J. | Hammerschlag, F.A.
Several factors were investigated for their influence on the transfer of an intron-containing beta-glucuronidase (GUS) gene into blueberry (Vaccinium spp.) leaf explants during the early stages of Agrobacterium-mediated gene transfer, including days of cocultivation, strain of Agrobacterium tumefaciens, explant age and genotype. The number of GUS-expressing leaf zones and calli were counted immediately and 2 weeks after cocultivation, respectively, to evaluate the gene transfer process. Agrobacterium tumefaciens strain EHA105 (pEHA105/p35SGUSint) was significantly more effective for transformation than strain LBA4404 (pAL4404/p35SGUSint). Four days of cocultivation with A. tumefaciens strain EHA105 yielded about 50-fold more GUS-expressing zones than 2 days of cocultivation. Significant differences among cultivars were observed for both GUS-expressing leaf zones and calli. For some cultivars, explant age influenced the number of GUS-expressing leaf zones and calli. In most cases, the number of GUS-expressing calli was highest in those cultivars where GUS expression in the leaves was high.
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