Optimization of Transformation Conditions of Potato by Agrobacterium tumefaciens and Introduction of AtNHX1 Gene

By mediation of Agrobacterium tumefaciens LBA4404,AtNHX1 gene was transferred into the microtubers of ‘Gannongshu 2’ potato.The key factors affecting the transformation were assessed and transgenic plants were generated and further tested using PCR-Southern blot hybridization.The results show that optimal kanamycin concentration for microtuber's differentiation and root's regeneration was 50 mg/L and acetosyringone had no impact on transformation of the microtubers.The optimized transformation conditions and procedures were as follows:microtubers were firstly immersed into liquid bacterium at OD600 of 0.5 for 8～10 min prior to pre-culture,followed by a two-day long co-culture before being transferred to and cultured in a differentiation medium for 40 days.A proportion 30% of the transgenic plants was kanamycin resistant.The PCR and PCR-Southern blot hybridization eventually detected the AtNHX1 gene inserted into the potato genome that could be used as raw materials for the development of salt resistant potato varieties.