DIY: Vacuum Agroinfiltration Using a Vacuum Desiccator and Vacuum Pump

Agrobacteria infiltration is a transient gene expression technique whereby Agrobacterium tumefaciens containing a gene circuit of interest are introduced (infiltrated) into plant leaf tissue either by injection or pulling a vacuum. Once the Agrobacterium are inside the leaf, transfer of the gene circuit from the Agrobacterium to leaf cells occurs and the gene circuit will then produce the protein/enzyme of interest in the plant. This technique is an alternative to using single cell (bacteria or mammalian) fermenters for protein production but can also be used for complementation studies (1), testing signal peptide localization (2), testing plant synthetic biology parts e.g. promoter testing (3) and systems analysis e.g. defense response (4). The transient nature of this technique allows relatively rapid testing and/or isolation of a protein/enzyme of interest (days) vs the time it would take to create a stable genetically modified plant (weeks to months).

In our poster we describe methodology and detail results for the implementation of whole leaf Agroinfiltration in Nicotiana benthamiana using a common laboratory vacuum desiccator and small vacuum pump.

1. Y. Shen et al., Virus Res 190, 25-33 (2014).
2. H. T. Parsons et al., Plant Cell 31, 2010-2034 (2019).
3. M. A. R. Sarkar, Plant Biotechnol (Tokyo) 36, 241-251 (2019).
4. A. S. M. Elnahal et al., Front Plant Sci 11, 919 (2020).