Short communication Assessment of transgenic maize events produced by particle bombardment or Agrobacterium-mediated transformation

Particle bombardment and Agrobacterium-mediated transformation are two popular methods currently used for producing transgenic maize. Agrobacterium-mediated transformation is expected to produce transformants carrying fewer copies of the transgene and a more predictable pattern of integration. These putative advantages, however, tradeoff with transformation efficiency in maize when a standard binary vector transformation system is used. Using Southern, northern, real-time PCR, and real-time RT-PCR techniques, we compared transgene copy numbers and RNA expression levels in R1 and R2 generations of transgenic maize events generated using the above two gene delivery methods. Our results demonstrated that the Agrobacterium-derived maize transformants have lower transgene copies, and higher and more stable gene expression than their bombardment-derived counterparts. In addition, we showed that more than 70% of transgenic events produced from Agrobacteriummediated transformation contained various lengths of the bacterial plasmid backbone DNA sequence, indicating that the Agrobacterium-mediated transformation was not as precise as previously perceived, using the current binary vector system. Abbreviations: ADH – alcohol dehydrogenase; bar – phosphinothricin acetyltransferase gene; CaMV 35S – Cauliflower Mosaic Virus 35S RNA Promoter; GOI – gene of interest; GUS – -glucuronidase; nos – nopaline synthase of Agrobacterium tumefaciens; NPK – Nicotiana protein kinase; PCR – Polymerase chain reaction; RT – Reverse transcription