Abstract
Transient expression systems are valuable tools in molecular biology. Agrobacterial infiltration of leaves is well-established in tobacco, but has led to limited success in the model plant Arabidopsis thaliana. An efficient expression system combining the advantages of Arabidopsis (well-characterised) and the simplicity of leaf infiltration is desirable. Here, I describe Agrobacterium tumefaciens-mediated transformation of Tropaeolum majus (nasturtium, order Brassicales) as a remarkably simple, cheap and highly efficient transient expression system. It provides the Arabidopsis community with a tool to study subcellular localisation, protein–protein interactions and reporter gene activities (e.g. luciferase, β-glucuronidase) in a genetic background that is closely related to their primary model organism. Unlike Arabidopsis , Tropaeolum is capable of engaging in endomycorrhizal associations and is therefore relevant also to symbiosis research. RNAi-based approaches are more likely to succeed than in the distantly-related Nicotiana transformation system. Tropaeolum majus was voted the “medicinal plant of the year 2013”. Conquering this plant for genetic manipulations harbours potential for biotechnological and pharmacological applications.
Highlights
The soil-born Agrobacterium tumefaciens is the only organism capable of interkingdom gene transfer
Plants of the order Brassicales produce characteristic sulfur-rich metabolites, glucosinolates. These mustard-oil glycosides function in the defence against pests and pathogens [19], and might be restrictive factor in Arabidopsis leaf transformation
This means that other species within the Brassicales may be accessible to Agrobacterium-mediated leaf infiltration
Summary
The soil-born Agrobacterium tumefaciens is the only organism capable of interkingdom gene transfer. It has been employed intensively for genetic manipulation of plant cells. Characterised transgenic individuals can be crossed, and individuals carrying both transgenes identified by genotyping. These approaches have their merits, substantial time and space are needed. Transgenes can be introduced into protoplasts obtained from suspension cultures or from freshly isolated leaf mesophyll cells. Easy transformation, allowing transfer of multiple transgenes at a time, has been achieved in Poinsettia [9].
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