Protocol for Defense Gene Silencing in Soybean Cotyledons

Roots initiated from cotyledons are transformed with Agrobacterium rhizogenes carrying an RNAi knockout construct for the gene of interest.  A GFP reporter gene is used to follow transformation.  Non-transformed roots are removed.

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Gene silencing introduced into the root moves systemically into the cotyledons.

 

 

    This image shows RNAi-based silencing of the GUS gene in

    cotyledons from a transgenic plant carrying an isoflavone

    synthase promoter-GUS fusion.  Cotyledons silenced for any

    gene of interest can next be used to examine responses to

    elicitors or infection.