Multiple T‑DNA Delivery to Plants Using Novel Mini Binary Vectors with Compatible Replication Origins

Improved plants are necessary to
meet human needs. Agrobacterium-mediated transformation
is the most common method used to rewire
plant capabilities. For plant gene delivery, DNA constructs are assembled
into binary T-DNA vectors that rely on broad host range origins for
bacterial replication. Here we present pLX vectors, a set of mini
binary T-DNA plasmids suitable for Type IIS restriction endonuclease-
and overlap-based assembly methods. pLX vectors include replicons
from compatible broad host range plasmids. Simultaneous usage of pBBR1-
and RK2-based pLX vectors in a two-plasmid/one-Agrobacterium strain strategy allowed multigene delivery to plants. Adoption of
pLX vectors will facilitate routine plant transformations and targeted
mutagenesis, as well as complex part and circuit characterization.