Workflow for a Functional Assay of Candidate Effectors From Phytopathogens Using a TMV-GFP-based System

Area of Science:

• Plant Pathology
• Molecular Biology
• Genetics

Background:

• Efficient screening of plant pathogen effectors is vital for crop disease resistance.
• Traditional methods are often slow and labor-intensive.
• A high-throughput assay is needed to accelerate the study of plant-pathogen interactions.

Purpose of the Study:

• To develop a robust, high-throughput screening assay for plant pathogen effectors.
• To enable rapid identification of effectors that modulate plant immunity.
• To provide a cost-effective and versatile tool for large-scale effector studies.

Main Methods:

• Utilized the tobacco mosaic virus-green fluorescent protein (TMV-GFP) vector system.
• Employed Agrobacterium-mediated transient expression in *Nicotiana benthamiana*.
• Monitored effector function via GFP fluorescence within six days using a UV lamp.

Main Results:

• Successfully established a high-throughput screening system for plant pathogen effectors.
• Demonstrated rapid evaluation of effector activity within six days.
• Showcased the system's ability to identify effectors interfering with plant immunity.

Conclusions:

• The TMV-GFP based system offers a significant advancement in screening plant pathogen effectors.
• This method accelerates the discovery of novel effectors and the development of disease-resistant crops.
• The assay is versatile, cost-effective, and applicable to various effector types and pathogens.