Plants mainly depend on both cell-surface and intracellular receptors to defend against various pathogens. The nucleotide-binding leucine-rich repeat (NLR) proteins are intracellular receptors that recognize pathogen effectors. The first NLR was cloned thirty years ago. Genomic sequencing and biotechnologies accelerated NLR gene isolation. NLR genes have been proven useful in breeding disease resistant crops. Here, we summarized the current knowledge of strategies for NLR gene isolation and provided a list of NLRs cloned in vegetables. We also discussed the mechanisms underlying NLR gene function, the challenges of NLRs in vegetable breeding and directions for future studies.