Verifying Plasmid Constructs via Transient Agrobacterium tumefaciens-Mediated Plant Transformation in Nicotiana...

Verifying plasmid constructs is a key, yet often overlooked step in plant molecular biology. Yet, due to lengthy biological cycles associated with many plant models and poor compatibility when extrapolating to other eukaryotic and prokaryotic models, it is important to test plant expression vectors before committing to the goal of obtaining stably transformed lines. Issues with vector expression can result in substantial time delays and the wasting of resources when not detected early on, especially when dealing with inducible vector expression systems as many inducible promoters present some degree of leakage.In this protocol, we present the detailed use of transient Agrobacterium tumefaciens transformation in Nicotiana benthamiana to confirm the expression of a gene of interest when expressed under an abiotic stress-inducible promoter.