NB-LRR class plant virus resistance gene is a one of the key players that shape the plant-virus interaction. Evolutionary arms race between plants and viruses often results in the breakdown of virus resistance in plants, which leads to a disastrous outcome in agricultural production. Although studies have analyzed the nature of plant virus resistance breakdown, it is still difficult to foresee the breakdown of a given virus resistance gene. In this chapter, we provide a protocol for evaluating the durability of plant virus resistance gene, which comprises the random mutagenesis of a virus gene, the introduction of the mutagenized gene into a virus context with highly efficient inoculation system, and the efficient screening of virus mutants that can overcome or escape a virus resistance.
Keywords: Agroinfection; Durability; Error-prone PCR; Infectious clone; Random mutagenesis; Resistance breakage.