Organogenesis and Sonication-Assisted Agrobacterium-Mediated Transformation of Poplar Roots

Abstract Functional genomics along with genetic transformation and plant regeneration are used to identify genes of interest, comprising essential tools to obtain plants with desired characteristics. In this study, we described an organogenesis and a genetic transformation protocol for Populus tremula x Populus alba clone 717-1B4 using roots. In the organogenesis experiments the PGRs zeatin, BA and kinetin at different concentrations were evaluated. The effect of explant age was evaluated and at 30 days “old” roots (6 months old) showed a higher regeneration rate when compared to “young” roots (2 months old). In the genetic transformation experiments kanamycin concentration, the use of sonication (SAAT), co-cultivation period and explant age were evaluated. Sonication positively affected transformation, while co-cultivation time did not interfere. Regarding explant age, no statistical differences were observed. Quantitative real-time PCR (qPCR) data showed that among the six transgenic plants evaluated, two presented two copies and the others only one copy of the gene. In this study, efficient protocols of organogenesis and genetic transformation for poplar roots are presented with a transformation efficiency of 58%.