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

ACS Synth Biol. 2017 Oct 20;6(10):1962-1968. doi: 10.1021/acssynbio.6b00354. Epub 2017 Jun 28.

Abstract

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.

Keywords: Agrobacterium tumefaciens; Potyviridae; Ugandan cassava brown streak virus; binary T-DNA vector; ethanol-inducible switch; multigene transfer; pBBR1; plant biotechnology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotechnology
  • DNA, Bacterial / genetics*
  • Genetic Vectors / genetics
  • Plants, Genetically Modified / genetics
  • Plasmids / genetics
  • Replication Origin / genetics*
  • Transformation, Genetic / genetics

Substances

  • DNA, Bacterial
  • T-DNA