Application of TILLING as a Reverse Genetics Tool to Discover Mutation in Plants Genomes for Crop Improvement

Abstract

The deployment of mutant resources for plant functional genomics is ever increasing with the availability of genome information of economically important crops. Targeting Induced Local Lesions IN Genomes (TILLING) is a reverse genetics approach for high-throughput discovery of induced mutations in the desired gene(s) from a mutant population developed through mutagenesis. TILLING accelerates trait discovery and crop genetic improvement. TILLING strategies for several crops have been developed for the identification of the desired mutants. The combined use of TILLING and the high-throughput sequencing technologies can overcome the bottlenecks of traditional TILLING approaches and facilitates the rapid discovery of useful mutations. Several important mutants related to economically important traits have been identified in agronomic and horticultural crops by employing TILLING by sequencing (TbyS) and, thus, have sizeable potential in crop improvement. In this chapter, we highlight significant developments in the use of TILLING strategies along with the application of high-throughput techniques, such as high-resolution melting (HRM) and next-generation sequencing (NGS) in the discovery of mutations and further discuss the challenges and prospects of TILLING in modern crop breeding.