T-DNA orientation, distance between two T-DNAs, and the transformation target cells significantly impact vector backbone integration and efficiency of generating marker-free transgenic plants in a co-transformation system.

ABSTRACT: The development of marker-free transgenic plants is essential to address biosafety concerns and facilitate regulatory approval. Co-transformation strategies involving separate T-DNAs for the gene of interest and selectable marker gene offer a clean approach but are often hampered by linked integration and vector backbone incorporation. In this study, we designed and evaluated a series of double T-DNA vectors with varying intervening sequence lengths and orientations to determine their impact on co-transformation efficiency and integration patterns in different plant species. Our results showed that shorter spacer regions increased the likelihood of linked T-DNA integration, while an ~3 kb intervening region minimized this risk. Contrary to previous findings, inverse orientation of T-DNAs w

SUBMITTER: Al Nuaimi M 

PROVIDER: S-EPMC12503609 | biostudies-literature | 2025 Oct

REPOSITORIES: biostudies-literature