SCREENING MUTATIONS IN DROUGHT TOLERANCE GENES USING SIMPLE METHOD FOR TILLING IN DURUM WHEAT (Triticum turgidum L. subsp. durum (Desf.))

Durum wheat (Triticum turgidum L. subsp. durum (Desf.) is an important crop in the Mediterranean Region, which requires meeting the demand for quality and quantity of this cereal. Induced mutation and single nucleotide polymorphism (SNP) detection technologies represent a modern challenge for identifying and manipulating genes for wheat improvement. The TILLING strategy makes use of traditional chemical mutagenesis followed by screening for single base mismatches to identify novel mutant loci. High-throughput TILLING (Targeting Induced local Lesions IN Genomes) methodology for screen mutations in mutagenized populations. Although TILLING has been combined to several pre-screening methods for SNP analysis which rely on expensive equipment.

This article focused how to make the technique TILLING more accessible and low cost and time saving on conventional agarose gels. A new durum wheat TILLING population has been developed by using 0.6 % ethyl methane sulfonate (EMS). To examine the efficiency of the mutagenic treatments, a pilot screening was carried out on 1500 mutants lines focusing on fives target genes related to drought tolerance (DHN11; pTd27; DHNWWZ; RUBISCO; SNAC).

Results from the first screens indicate 31 novel alleles identified in 5 targeted genes from TILLING population of durum wheat. That the mutagen treatment was effective with an average mutation of at least one mutation per 19-47 kb.

This work illustrated the applicability and efficiency of a simple method by using a non-denaturing agarose gel system. This detection of induced variability in durum wheat TILLING population useful for a better understanding of key gene functions.