A novel efficient method for maize genetic transformation: Usage of agrobacterial monolayer

摘要

A new approach for maize genetic transformation using Agrobacterium tumefaciens was developed. It allows the usage of both immature embryos and embryogenic calluses as explants. This method includes long-term (15-20 days) cultivation of maize tissues on the agrobacterial monolayer. In this case, necrotic lesions did not appear and all cells were alive. Regenerated plants expressed genes for neomycine phosphotransferase and beta-glucuronidase. The presence of the nptII gene was confirmed by PCR analysis and Southern-blot hybridization. In dependence on maize genotype and experimental conditions, the frequency of genetic transformation varied from 1.5 to 16%. Inheritance of genes introduced was demonstrated. The method elaborated could be applied to a wide range of maize genotypes.