ABSTRACT
The rapid advancement of large-scale genome sequencing projects has opened new avenues for the application of conventional mutation techniques in not only forward but also in reverse genetics strategies. Targeting induced local lesions in genomes (TILLING), developed as an alternative to cumbersome insertional mutagenesis, takes advantage of classical mutagenesis, sequence availability and high throughput screening for nucleotide polymorphisms in a targeted sequence. Since it is readily applicable to most plants, it remains a dominant non-genetic manipulation (GM) method for obtaining desirable mutations in known genes. The application of next-generation sequencing (NGS) has been a breakthrough change, which permits multiplexing of gene targets and genomes. In this chapter, we review here the discovery and technology with more focus on the ease of applications, Ecotilling, constraints, recent developments in polyploids crops, and the future of TILLING for crop breeding.
