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ARS Home » Plains Area » Fargo, North Dakota » Edward T. Schafer Agricultural Research Center » Cereal Crops Improvement Research » Research » Research Project #440208

Research Project: Functional Characterization of Pathogen Produced Effectors and Their Host Targets

Location: Cereal Crops Improvement Research

Project Number: 3060-22000-051-003-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Jun 1, 2021
End Date: May 31, 2026

Objective:
The objectives of this cooperative research project are to: 1) Functionally characterize SnTox267 by identifying critical amino acids involved in targeting Snn2, Snn6, and Snn7. 2) Identify host target inter-actors for SnTox267. 3) Identify SnTox5 amino acids critical for inducing necrosis and colonizing the leaf mesophyll.

Approach:
The Parastagonospora nodorum-wheat interaction involves a diverse set of pathogen-produced effectors that target host gene products to gain entry and/or colonize wheat. Recently we cloned and validated two new pathogen produced necrotrophic effectors including SnTox5 and SnTox267. SnTox5 is unique in that it has been shown to target the wheat gene Snn5 to induce programmed cell death resulting in the availability of host cell nutrient necessary for the pathogen to complete its pathogenic life cycle. In addition to the targeting of Snn5, SnTox5 was also shown to be critical to the colonization of the mesophyll layer of the wheat leaf even in the absence of Snn5. SnTox267 is a unique effector in that this protein targets two different host pathways to induce necrosis, one involving Snn2/Snn6 and the other involving Snn7. The grad student working on this project will use CRISPR-based gene editing to identify the amino acids involved in the interaction with Snn5 that results in cell death as well as the amino acids important in the colonization of the mesophyll layer. Additionally the student will use CRISPR-based gene editing of SnTox267 to functionally characterize how the pathogen is using this protein to target multiple host pathways. Yeast two-hybrid will also be used to identify direct protein-protein interactions involving SnTox5 and SnTox267.