Targeting Induced Local Lesions in Genomes (TILLING): advances and opportunities for fast tracking crop breeding

Abstract

The intensification of food production via conventional crop breeding alone is inadequate to cater for global hunger. The development of precise and expeditious high throughput reverse genetics approaches has hugely benefited modern plant breeding programs. Targeting Induced Local Lesions in Genomes (TILLING) is one such reverse genetics approach which employs chemical/physical mutagenesis to create new genetic sources and identifies superior/novel alleles. Owing to technical limitations and sectional applicability of the original TILLING protocol, it has been timely modified. Successions include: EcoTILLING, Double stranded EcoTILLING (DEcoTILLING), Self-EcoTILLING, Individualized TILLING (iTILLING), Deletion-TILLING (De-TILLING), PolyTILLING, and VeggieTILLING. This has widened its application to a variety of crops and needs. They can characterize mutations in coding as well as non-coding regions and can overcome complexities associated with the large genomes. Combining next generation sequencing tools with the existing TILLING protocols has enabled screening of huge germplasm collections and mutant populations for the target genes. In silico TILLING platforms have transformed TILLING into an exciting breeding approach. The present review outlines these multifarious TILLING modifications for precise mutation detection and their application in advance breeding programmes together with relevant case studies. Appropriate use of these protocols will open up new avenues for crop improvement in the twenty first century.

Keywords:

• ComSeq
• DEcoTILLING
• exome capture
• mutation breeding
• TILLING modifications
• TILLING platforms

Acknowledgments

Authors thank Director, Indian Council of Agricultural Research (ICAR)-Indian Agricultural Research Institute (IARI), New Delhi, Head, Division of Genetics, ICAR- IARI, New Delhi for healthy discussions during preparation of this article.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Additional information

Funding

This work has been financially supported by grant received from Indian Agricultural Research Institute, New Delhi (Project no- JAN 09/16). The funding bodies had no role in the design of the review and writing the manuscript.