Transformation of androgenic-derived Festulolium plants (Lolium perenne L. x Festuca pratensis Huds.) by Agrobacterium tumefaciens

Genetic transformation of androgenic-derived amphidiploid Festulolium plants (Lolium perenne L. x Festuca pratensis Huds., 2n = 4x = 28) by Agrobacterium tumefaciens has been achieved. Anther culture-induced calli of Festulolium Bx351 were inoculated with Agrobacterium tumefaciens strain LBA4404 carrying pIG121-Hm encoding the hygromycin resistance (hph) and beta-glucuronidase (uidA) genes under the control of a CaMV 35S promoter. Twenty-three putative transformants were obtained from the hygromycin selection, 19 of which (82.6%) showed GUS activity. The integration of transgene was detected by using genomic DNA PCR analysis, RT-PCR analysis and Southern blot hybridization, respectively, which revealed that foreign gene was integrated into the genomes of dihaploid transformants (2n = 2x = 14). The haploid embryogenic system offers a stable means of transformation, as the introduced trait can be readily fixed through chromosome doubling.