Agrobacterium-mediated transformation of Solanum phureja

A population of transgenic plants was produced by the transformation of internodal explants of Solanum phureja, DB337/37 ( the cultivar Mayan Gold) using an Agrobacterium tumefaciens LBA4404-based vector containing a phytoene synthase gene ( crtB). The regeneration strategy utilised a two-step protocol, with a 12-day callus induction stage mediated by 1.07 mu M alpha-napthaleneacetic acid (NAA), 7.10 mu M zeatin riboside and 0.06 mu M gibberellic acid (GA3), followed by a prolonged ( up to 90 day) shoot induction stage on medium containing 0.11 mu M NAA, 7.10 mu M zeatin riboside and 0.06 mu M GA3 supplemented with kanamycin at 50 mg l(-1) as the selection agent. Southern analysis of the transgenic population revealed that the transgene copy number varied between one and five in the lines tested. Northern blot analysis showed significant expression of the introduced crtB gene in some lines during tuber development. Cytological analysis of the material showed a high incidence of chromosome doubling in the transgenic population with over 80% of all lines tested having doubled their chromosome complement during the transformation process.