Stability of inheritance of transgenes in maize (Zea mays L.) lines produced using different transformation methods

Abstract

The inheritance and stability of the acetolactate synthase (als) transgene were compared in transgenic maize plants, generated using the pollen-tube pathway, particle bombardment, or Agrobacterium-mediated methods of transformation. Progeny populations generated by successive selfing or backcrossing of primary transformants were analyzed over three generations, using PCR and herbicide screening, to examine segregation and als activity, respectively, and transgenic homozygous plants were selected. The pollen-tube method resulted in a higher rate of primary normal transgenic plants and a less-stable transmission of the als locus than did the other two methods. When transferred by the particle bombardment and Agrobacterium-mediated methods, the als gene was in a much higher proportion of Mendelian transmission than transferred by the pollen-tube method. Compared to the Agrobacterium-mediated transformation, the particle bombardment method tends to create multiple copies and insert sites of the als gene in maize genome, which delaying the homogenization of the als locus with advancing generations. Agrobacterium-mediated transformation resulted in a greater proportion of stable, low copy number (in general 1–2) transgenic events, facilitating the stable inheritance of the als gene, and producing multiple desirable transgenic plants.