Agrobacteriumbased

Agrobacterium-based transformation refers to techniques that use the bacterium Agrobacterium tumefaciens or related species to introduce foreign DNA into plant cells. The natural system centers on a Ti (tumor-inducing) plasmid that transfers a defined region called T-DNA into the plant genome, a process that causes crown gall disease. Researchers repurpose this mechanism to deliver genes of interest and produce stable, transgenic plants under appropriate biosafety controls.

Most widely used is the binary vector system, where a disarmed Ti plasmid supplies the virulence functions and a separate plasmid carries the gene of interest flanked by T-DNA borders. During infection, Vir genes orchestrate transfer; VirD2 nicks the T-DNA borders and VirE2 coats the DNA, guiding it into plant cells and enabling integration into the genome via the plant’s repair machinery. The result is typically a heritable insertion of the transgene.

Applications encompass a broad range of plant biotechnology goals, including crop traits such as pest resistance, herbicide tolerance, and enhanced nutrition, as well as functional genomics studies. The method is also used for transient expression in tissues via agroinfiltration and, with ongoing optimization, for transforming a wider array of species. Advances have extended use to genome-editing approaches that deliver CRISPR components through Agrobacterium.

Advantages include relatively high transformation efficiency and a propensity for stable, single- or low-copy insertions with predictable expression in many species. Limitations involve species- and cultivar-specific host range, potential insertional effects on endogenous genes, and regulatory or public acceptance considerations. Safety is generally favorable when using disarmed strains within established biosafety frameworks.