Figure 5

Phenotype of MT and MT- Rg1 and selection of transgenic lines. A. Phenotype of adult MT and MT-Rg1 plants. The presence of the r and Rg1 alleles resulted in yellow fruits and branched shoot, respectively, in the MT-Rg1 genotype (right). B. In vitro regeneration in absence (top) and presence (bottom) of 100 mg/L kanamycin. The two plates at the bottom of the figure contain explants pre-incubated with Agrobacterium harboring the nptII-containing vector pROKII. C. Analyses of nptII gene in acclimated T₀ transgenic plants by agarose gel electrophoresis of PCR amplification of a 700 bp fragment of nptII gene and DNA molecular marker (100 bp). D. Selection of segregating T₁ lines performed in the greenhouse by spraying 400 mg/L kanamycin in 14-day-old seedlings for 5 consecutive days. E. Analysis of nptII expression by qRT-PCR of T2 single copy homozygous plants transformed with the pROKII vector containing the nptII and AtCKX2 genes. F. GUS staining of leaf disks excised from acclimated T₀ plants transformed with the pGPTV-GUS-KAN containing the gusA gene under control of the wound inducible JERE element (AGACCGCC). Note the enhanced staining of the disk borders, which correspond to wounded areas. Bars = 4 cm (A, B and D) and 1 mm (F). Non-transformed MT plants were used as controls (E, F).