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Effect of initial explant on effectivity of Agrobacterium tumefaciens-mediated transformation of Gerbera hybrida

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Nowak E., Rojek Z., Kucharska D., Orlikowska T.
Biotechnologia
|
1997
|
issue 4
27-37
EN
An efficient method for genetic Agrobacterium tumefaciens - mediated transformation of five cultivars Gerbera hybrida was established. The youngest leaves and shoot tips from proliferating in vitro cultures were co-cultivated with disarmed strain LBA 4404/pBI121 carrying the chimaeric genes nptII and gus. The inoculated explants were repeatedly cultured on regeneration medium with kanamycin and Biotaxym. After 9-10 months from 0.04 to 0.25 independently transformed shoots from one inoculated leaf and from 0.06 to 0.65 from one inoculated shoot were obtained, depending on cultivar and additional treatment. Transformed shoots accounted for 5.0 to 50.0% of all regenerated shoots. Additional wounding` of leaves before inoculation by squeezing and scratching off increased transformation efficiency. Better results for recalcitrant to regeneration cultivars were obtained if squeezed shoot tips were inoculated. Regenerating shoots were selected on kanamycin, screened for gus expression with X-gluc and tested by PCR for nptII gene. All together 162 transgenic shoots were obtained in cvs Amber, Boy, Ferrari, Mariola and Tamara, but only 24 relatively stable lines were established.
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