Is it the end of TILLING era in plant science?

Abstract

Since its introduction in 2000, the TILLING strategy has been widely used in plant research to create novel genetic diversity. TILLING is based on chemical or physical mutagenesis followed by the rapid identification of mutations within genes of interest. TILLING mutants may be used for functional analysis of genes and being nontransgenic, they may be directly used in pre-breeding programs. Nevertheless, classical mutagenesis is a random process, giving rise to mutations all over the genome. Therefore TILLING mutants carry background mutations, some of which may affect the phenotype and should be eliminated, which is often time-consuming. Recently, new strategies of targeted genome editing, including CRISPR/Cas9-based methods, have been developed and optimized for many plant species. These methods precisely target only genes of interest and produce very few off-targets. Thus, the question arises: is it the end of TILLING era in plant studies? In this review, we recap the basics of the TILLING strategy, summarize the current status of plant TILLING research and present recent TILLING achievements. Based on these reports, we conclude that TILLING still plays an important role in plant research as a valuable tool for generating genetic variation for genomics and breeding projects.

Phenotypic and genetic characterization of an Avena sativa L. germplasm collection of diverse origin: implications for food-oat breeding in Chile

“…At least two thousand rice, barley, wheat, soybean, maize, and oat mutant varieties have been released to farmers ( https://nucleus.iaea.org/sites/mvd/SitePages/Home.aspx ). The development of Targeting Induced Local Lesion IN Genomes (TILLING), consisting in mutagenesis followed by a rapid identification of mutations in the genes of interest ( Szurman-Zubrzycka et al., 2023 ), has been broadly used to improve traits such as starch synthesis, plant architecture, disease resistance, drought and salinity tolerance, and other yield parameters in cereals ( Irshad et al., 2020 ; Nouman-Khalid et al., 2021 ; Abdelnour-Esquivel et al., 2010 ). In oats, TILLING allowed to induce variation in genes encoding for enzymes involved in the pathways of lignin and beta-glucan biosynthesis ( Chawade et al., 2010 ).…”