Effect of Explant, Genotype and Plant Growth Regulators on Regeneration and Agrobacterium -Mediated Transformation of Potato

摘要

The objective of the present study is to develop an efficient protocol for plant regeneration and Agrobacterium -mediated transformation of potato cultivars Desiree, Agria and Marfona grown under Iranian agricultural conditions. The regeneration efficiency from internode, leaf and petiole explants was examined. The results from all Shoot Induction Medium (SIM) combinations and all cultivars together indicate highly efficient shoot regeneration from internode (4.56 shoots per explant). The response of petiole and leaf explants was lower, 3.83 and 2.55 shoots per explant, respectively. The highest efficiency of shoot regeneration was achieved with internode explants of cultivar Desiree (6.64 shoots per explant) on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ) 1 mg L^?1 + 6-benzyladenine (BAP) 1 mg L^?1. For plant transformation, internode explants were inoculated and co-cultivated with Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pBI-121 containing β-glucuronidase (GUS) and nptII genes. Reverse transcriptase- polymerase chain reaction analysis and histochemical assay for β-glucuronidase indicated that the gene coding for this enzyme was integrated in the potato genome and could be expressed in potato tissues. The presence of nptII gene in the kanamycin resistant plants was verified by polymerase chain reaction analysis. The transformation frequency ranged from 22-42%.