[Research on Festuca arundinacea transformation mediated by Agrobacterium tumefaciens].

Abstract

The embryo-derived calli from four types of tall fescues (Festuca arundinacea Schreb) were transformed with two Agrobactrium tumefaciens strains AGL1 and GV3101. AGL1 harbors an intron-AtNHX1 expression vector pROK2U containing ubiqutin promoter and npt II marker gene. GV3101 harbors an intron-AtNHX1 expression vector pROK2 containing 35S promoter and npt II gene. After infection and co-culture with AGL1 or GV3101, the embyogenic calli were selected with 50-150 mg/L paromomycine and 1126 resistant plants regenerated from the resistant calli. All plants were selected further with 10-20 mg/L Kanamycin and 525 of them remained green. Genome DNA of the resistant plants was checked with specific primers and probe from AtNHX1 gene. The results of PCR assay and Southern blot analysis indicted that exogenous target gene (AtNHX1 gene) had been transferred into different cultivars of Festuca arundinacea. Different transformation frequencies among the four cultivars were obtained.