Transformation of potato (Solanum tuberosum l.) using tuber discs and stem explants

Abstract

Transformation of three potato cultivars (Desiree, Isola and Anag) was achieved using tuber discs and callus tissue derived from cultured stems via co-cultivation with Agrobacterium tumafaciens Ti plasmid based vectors. Neomycin phosphotransferase (NPTII) marker and (3-glucuronidase (GUS) reporter genes were used in transfer-optimization studies. First selection of transgenics was done on selective Murashige and Skoog (MS) media containing 100 mg/l kanamycin and 500 mg/l cefotaxime. Histochemical assays were done for GUS activity using X-Gluc substrat. In addition polymerase chain reaction based Southern blot analyses revealed the existence of the transferred genes. The highest transformation frequency (10%) was obtained with cultivar Desiree in MS media supplied with 5 mg/l zeatin riboside and 1.5 mg/l indolacetic acid. However, A nag gave the lowest frequency using tuber discs (3%) and stem explants (2%). This protocol could be useful for the improvement ofpotato through gene manipulation. © 1995 Taylor & Francis Group, LLC.