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Updated: May 15, 2026

VIGS-Mediated Forward Genetics Screening for Identification of Genes Involved in Nonhost Resistance

VIGS-Mediated Forward Genetics Screening for Identification of Genes Involved in Nonhost Resistance

Published on: August 23, 2013

VIGS-Based Screening for NLR Genes Regulating Viral Effector-Induced Hypersensitive Response.

Wenqi Mao¹, Jianping Chen¹, Fei Yan²

¹State Key Laboratory for Quality and Safety of Agro-products, Key Laboratory of Biotechnology in Plant Protection of MARA, Key Laboratory of Green Plant Protection of Zhejiang Province, Institute of Plant Virology, Ningbo University, Ningbo, China.

Methods in Molecular Biology (Clifton, N.J.)

|May 13, 2026

Summary

This study presents a fast method to identify plant immune genes. Using virus-induced gene silencing (VIGS) in Nicotiana benthamiana, researchers can quickly find nucleotide-binding leucine-rich repeat (NLR) genes involved in plant defense against pathogens.

Keywords:

Hypersensitive response Nucleotide-binding leucine-rich repeat protein Tobacco rattle virus Virus-induced gene silencing

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Published on: March 2, 2014

Area of Science:

Plant immunity
Molecular genetics
Biotechnology

Background:

Nucleotide-binding leucine-rich repeat (NLR) proteins are crucial for plant defense against pathogens, recognizing specific effector proteins and triggering immune responses.
Improving crop resistance by utilizing NLR genes is economically vital, but traditional identification methods are slow and labor-intensive.
RNA interference (RNAi) offers sequence-specific gene silencing, providing a potential avenue for efficient gene discovery.

Purpose of the Study:

To develop and detail a rapid screening protocol for identifying host NLR genes involved in viral effector-activated hypersensitive response (HR).
To demonstrate the utility of this protocol using a specific viral effector, turnip mosaic virus-coded nuclear inclusion protein a protease (NIa-Pro), in Nicotiana benthamiana.

Main Methods:

Utilized a tobacco rattle virus-mediated virus-induced gene silencing (VIGS) platform in Nicotiana benthamiana.
Employed VIGS for high-throughput screening to identify NLR genes mediating HR against viral effectors.
Focused on the NIa-Pro effector from turnip mosaic virus as a case study.

Main Results:

Successfully established a VIGS-based screening platform for rapid identification of plant NLR genes.
Demonstrated the platform's effectiveness in identifying host NLR genes involved in the HR response triggered by the NIa-Pro effector.
The protocol facilitates faster discovery compared to traditional methods.

Conclusions:

Virus-induced gene silencing (VIGS) in Nicotiana benthamiana is an efficient strategy for fast identification of host NLR genes involved in plant immunity.
This method accelerates the discovery of resistance genes, aiding in the development of improved crop varieties.
The developed protocol offers a valuable tool for plant-pathogen interaction research.