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ARS Home » Midwest Area » Ames, Iowa » Corn Insects and Crop Genetics Research » Research » Research Project #432417

Research Project: Identification and Characterization of Host Targets of Powdery Mildew Effector Proteins

Location: Corn Insects and Crop Genetics Research

Project Number: 5030-21000-067-01-I
Project Type: Interagency Reimbursable Agreement

Start Date: Feb 1, 2017
End Date: Jan 31, 2020

Objective:
The primary focus of this research is to identify host targets of Blumeria effector proteins and characterize their role(s) in barley immune responses to this important fungal pathogen. Objective 1 will identify barley protein targets of the selected Blumeria effectors using a novel strategy to identify and rank candidate interacting proteins. Objective 2 will investigate global changes in host and pathogen proteomes over the first 48 hours of Blumeria infection. Objective 3 will integrate results from Objectives 1 and 2 to identify and functionally characterize barley proteins that contribute to disease resistance and/or are exploited by Blumeria to facilitate disease progression.

Approach:
Existing gene expression data have been used to identify Blumeria effectors that are expressed at distinct stages of the infection process and differentially expressed among resistant and susceptible host genotypes. Objective 1 will utilize a novel yeast two-hybrid approach coupled with next-generation sequencing that will expedite the identification and ranking of candidate interacting proteins. Objective 2 will use high resolution mass spectrometry to investigate global changes in the proteomes of both pathogen and host during over the first 48 hours of infection in both resistant and susceptible genotypes. Objective 3 will use various clustering and network analysis techniques to identify high priority protein-protein interactions for further study. Functional analysis of these interactions will include disease phenotyping, localization, and enzymatic assays to determine the role(s) of specific barley and Blumeria proteins in powdery mildew disease resistance/susceptibility.