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A diploid wheat TILLING resource for wheat functional genomics

DOI: 10.1186/1471-2229-12-205

Keywords: TILLING, Reverse genetics, Triticum monococcum, Mutation frequency, Waxy, Lignin

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Abstract:

We developed a TILLING population of 1,532 M2 families using EMS as a mutagen. A total of 67 mutants were obtained for the four genes studied. Waxy gene mutation frequencies are known to be 1/17.6 - 34.4 kb DNA in polyploid wheat TILLING populations. The T. monococcum diploid wheat TILLING population had a mutation frequency of 1/90 kb for the same gene. Lignin biosynthesis pathway genes- COMT1, HCT2, and 4CL1 had mutation frequencies of 1/86 kb, 1/92 kb and 1/100 kb, respectively. The overall mutation frequency of the diploid wheat TILLING population was 1/92 kb.The mutation frequency of a diploid wheat TILLING population was found to be higher than that reported for other diploid grasses. The rate, however, is lower than tetraploid and hexaploid wheat TILLING populations because of the higher tolerance of polyploids to mutations. Unlike polyploid wheat, most mutants in diploid wheat have a phenotype amenable to forward and reverse genetic analysis and establish diploid wheat as an attractive model to study gene function in wheat. We estimate that a TILLING population of 5, 520 will be needed to get a non-sense mutation for every wheat gene of interest with 95% probability.Common or bread wheat Triticum aestivum L. is an allohexaploid having three genomes A, B and D and a huge genome size of ~17 Gb [1,2]. For gene functional analysis, all the three homoeologous loci have to be individually targeted and subsequently combined to evaluate the phenotypes. This may take years. Although this is necessary for many genes of agronomic interest, it is cumbersome for other genes where the biological function has not been validated in wheat. Triticum monococcum is a cultivated A-genome diploid wheat and can be used to study traits, genes, and alleles as a model for bread wheat. Establishing phenotype to genotype relationships and allocating function of variant alleles in diploid wheat is considerably straightforward. As an alternative to studying gene functions in hexaploid wh

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