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

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Abstract
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 β-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.

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