Transformation of P_(SAG12)-ipt Gomphosis Gene into Potato

A population of transgenic plants was produced by the transformation of internodal explants of potato using an Agrobacterium tumefaciens LBA4404-based vector containing a gomphosis gene(PSAG12-ipt).The results showed that intermodal explants were more effective for transformation than leaf explants.The regeneration strategy utilised a three-step protocol,a 2 days pre-culture on the MS medium containing 6-benzylaminopurine(6-BA) 0.25 mg/L,α-naphythy lacetic acid(NAA) 0.25 mg/L,2,4-dichlorophenacetic acid(2,4-D) 0.25 mg/L,and supplemented 1% Na2SO3,followed the explants of pre-culture were incubated 8 min in Agrobacterium tumefaciens LBA4404 suspension(OD600 0.2~0.5),last,the explants were co-cultured 3 days on basal medium without supplemented any phytohormones.After 3 days co-cultivation,the explants of internode and leaf were transferred to basal medium supplemented phytohormones,1% Na2SO3,200 mg/L cefotaxime and 75 mg/L kanamycin until to regenerated plants.Transgenic plants were identified utilizing PSAG12 and ipt gene dual primer by PCR.The positive transformation rate was 65.8%.Southern blotting analysis identification showed that most ipt gene were induced into potato genome only one copy.