A functional genomics method for assaying gene function in phytopathogenic fungi through host-induced gene silencing mediated by agroinfiltration.
With the rapid growth of genomic information, there is an increasing demand for efficient analysis tools to study the function of predicted genes coded in genomes. Agroinfiltration, the delivery of gene constructs into plant cells by Agrobacterium tumefaciens infiltrated into leaves, is one such versatile, simple, and rapid technique that is increasingly used for transient gene expression assay in plants. In this chapter, we focus on the use of agroinfiltration as a functional genomics research tool in molecular plant pathology. Specifically, we describe in detail its use in expressing phytopathogenic fungal gene sequences in a host plant to induce RNA silencing of corresponding genes inside the pathogen, a method which has been termed host-induced gene silencing (HIGS). We target the fungal pathogen Puccinia triticina which causes leaf rust on its wheat host, but the method is applicable to a variety of pathosystems.
Pacific Agri-Food Research Centre, Agriculture and Agri-Food Canada, PO Box 5000, 4200 Hwy 97, Summerland, BC, Canada, V0H 1Z0.
Vinay
V
Panwar
Brent
B
McCallum
Guus
G
Bakkeren
D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000502:physiology*; D001487:Basidiomycota / Q000502:physiology*; D005656:Fungal Proteins / Q000235:genetics*; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D005822:Genetic Vectors / Q000235:genetics; D023281:Genomics / Q000379:methods*; D054884:Host-Pathogen Interactions; D010935:Plant Diseases / Q000382:microbiology; D018515:Plant Leaves / Q000382:microbiology; D011994:Recombinant Proteins / Q000235:genetics; D014908:Triticum / Q000382:microbiology*
D005656:Fungal Proteins; D011994:Recombinant Proteins