Genetic transformation of potato (Solanum tuberasum L.) to produce resistant plants for potato beetle (Leptinotarsa decemlineata Say.)

Two different EHA 101 isolates of Agrobacterium tumefaciens harbouring pCAMBIA1301 and PGA482GG plasmids were used for the genetic transformation of two potato varieties. The plasmid, pCAMBIA 1301 carrying Hpt gene provides resistance to antibiotic of hygromycin B, and Cry 1A(c) gene provides resistance to some insects. The plasmid, pGA482GG carrying Npt II provides resistance to antibiotic of kanamycin and GUS provides visible transformants. Genetic transformation was conducted on leaf and tissue fragments of Marfona and Granola Potato (Solanum tuberasum L.) varieties. The transgenic patato plants contaning Npt II and GUS genes in 50 mg/l kanamycin and transgenic outgrowth contaning Cry 1A(c) gene in 10 mg/l hygromycin B, were selected in L medium. ? -glucuronidase activity in plants contaning Npt II and GUS genes were determined by the analysis of histochemical GUS assay. The PCR process conducted on ten plant with GUS positive resulted in nine plant samples generating specific 660 bp DNA band. The PCR process for Npt II gene resulted in seven positives out of eleven samples. Thirty-five plants out of fifty-five plants resistant to hygromycin B antibiotic, were determined by specific 700 bp DNA fragment reproduced with PCR analysis to have Hpt gene. In the five transgenic microtubers of Marfona variety was also determined to have Hpt gene. Transgenic Marfona and Granola potato varieties having Hpt gene were biologically tested with potato beetles. Testing showed that % 80-85 of larvas of potato beetles lived on transgenic Marfona and Granola varieties died