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Traditional genetic methods for identifying disease-resistant nucleotide binding and leucine-rich repeat-containing receptors (NLRs) proteins are often laborious and time-consuming. To streamline the screening process, we employed a pre-constructed combinatorial Virus-Induced Gene Silencing (VIGS) library. This library targets 301 NbNLR genes by integrating specific fragments from 4 to 6 distinct NLRs into a single VIGS vector, generating 55 VIGS combination (VIGS-com). This chapter presents an efficient method based on tobacco rattle virus (TRV)-mediated VIGS for rapidly screening NLR genes that confer resistance to plant virus infection in Nicotiana benthamiana, using turnip mosaic virus (TuMV) as a case. This approach enables rapid and systematic screening of key disease-resistant NLR receptors in antiviral plant immunity.

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Virus-Induced NLR-Silencing Library-Based Screening to Identify Antiviral NLR Genes

  • 实验方案

  • 发表日期：

    2026年

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  • Jiajia Lin,
  • Jianping Chen,
  • Fei Yan,
  • Guanwei Wu

摘要

Traditional genetic methods for identifying disease-resistant nucleotide binding and leucine-rich repeat-containing receptors (NLRs) proteins are often laborious and time-consuming. To streamline the screening process, we employed a pre-constructed combinatorial Virus-Induced Gene Silencing (VIGS) library. This library targets 301 NbNLR genes by integrating specific fragments from 4 to 6 distinct NLRs into a single VIGS vector, generating 55 VIGS combination (VIGS-com). This chapter presents an efficient method based on tobacco rattle virus (TRV)-mediated VIGS for rapidly screening NLR genes that confer resistance to plant virus infection in Nicotiana benthamiana, using turnip mosaic virus (TuMV) as a case. This approach enables rapid and systematic screening of key disease-resistant NLR receptors in antiviral plant immunity.