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ARS Home » Plains Area » Fargo, North Dakota » Edward T. Schafer Agricultural Research Center » Cereal Crops Research » Research » Research Project #430238

Research Project: Cloning and Validation of New Parastagonospora nodorum Necrotrophic Effectors and Further Characterization of SnTox1 and the SnTox1-Snn1 Interaction

Location: Cereal Crops Research

Project Number: 3060-22000-050-33-I
Project Type: Interagency Reimbursable Agreement

Start Date: Feb 1, 2016
End Date: Jan 31, 2020

Objective:
We will use novel approaches to identify additional effector-host gene interactions. New necrotrophic effector (NE) candidates will be identified using proven approaches as well as novel approaches that will lead to the identification of NE interactions never before identified. We will also continue the investigation of the mode of action, localization, and specific interacting molecules of SnTox1. This analysis of the mode of action of SnTox1 will provide critical information as to how S. nodorum is causing disease.

Approach:
Currently three necrotrophic effector (NE) genes have been cloned and published and several more are being validated. It is the objective of this proposal to identify and characterize additional NE-host gene interactions in order to better understand not only this necrotrophic host pathogen system but also to provide valuable information about how necrotrophic specialists in general are modulating the host to induce disease. We have crudely identified several specific NE host gene interactions using both culture filtrates and fungal inoculations in conjunction with wheat mapping populations to genetically characterize this system. The NEs corresponding to these host genes will be characterized, purified, and the corresponding genes cloned and validated using methods proven to be effective in our lab including medium and high pressure liquid chromatography, 2-dimensional gel electrophoresis, and gene transformation, and heterologous expression. Strong preliminary genome wide association study (GWAS) data has located several candidate gene regions not previously identified. Genes in these regions will be prioritized and functionally characterized. Additionally, we will focus a deeper study on the SnTox1-Snn1 interaction, specifically looking at the mode of action of SnTox1. This will involve the study of SnTox1 localization and the identification of possible protein-protein interactions involving SnTox1. Mode of action studies will focus on site directed mutagenesis of protein sites thought to be involved in protein-protein communication as well as sites involved in chitin binding that are important in protection from wheat chitinases.