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Development of Targeting Induced Local Lesions IN Genomes (TILLING) Populations in Small Grain Crops by Ethyl Methanesulfonate Mutagenesis
Published on: July 16, 2019
A diploid wheat TILLING resource for wheat functional genomics.
Nidhi Rawat1, Sunish K Sehgal, Anupama Joshi
1Wheat Genetic and Genomic Resources Center, Throckmorton Hall, Kansas State University, Manhattan, KS 66506, USA.
BMC Plant Biology
|November 9, 2012
Summary
We developed a Triticum monococcum (einkorn wheat) TILLING population, revealing mutation frequencies suitable for gene analysis. This diploid wheat model offers advantages for studying gene function, despite lower mutation rates than polyploid wheat.
Area of Science:
- Plant genetics
- Crop domestication
- Functional genomics
Background:
- Triticum monococcum L. is the first domesticated diploid wheat, serving as a genetic model for wheat traits.
- Diploid wheat lacks amenability to reverse genetics methods like insertion mutagenesis.
- Targeting Induced Local Lesions IN Genomes (TILLING) provides a functional genetics approach for wheat gene analysis.
Purpose of the Study:
- To develop a TILLING population in diploid wheat (Triticum monococcum).
- To determine mutation frequencies for specific genes in this diploid TILLING population.
- To evaluate the utility of diploid wheat as a model for functional genetics.
Main Methods:
- Developed a TILLING population of 1,532 M2 families using ethyl methanesulfonate (EMS) as a mutagen.
- Analyzed mutation frequencies for the Waxy gene and lignin biosynthesis pathway genes (COMT1, HCT2, 4CL1).
- Calculated overall mutation frequency within the diploid wheat TILLING population.
Main Results:
- Obtained 67 mutants across four genes studied.
- Observed mutation frequencies of 1/90 kb for the Waxy gene in diploid wheat.
- Lignin biosynthesis genes showed frequencies of 1/86 kb (COMT1), 1/92 kb (HCT2), and 1/100 kb (4CL1).
- Overall mutation frequency in the diploid T. monococcum TILLING population was 1/92 kb.
Conclusions:
- The diploid wheat TILLING population exhibits higher mutation frequency than other diploid grasses but lower than polyploid wheat.
- Mutants in diploid wheat generally display phenotypes amenable to genetic analysis.
- Diploid wheat is established as a valuable model for studying gene function in wheat.
- A TILLING population of 5,520 is estimated to be necessary for a 95% probability of obtaining a nonsense mutation for any gene of interest.
