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United States Department of Agriculture

Agricultural Research Service

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Research Project: Development and Characterization of a Durum Wheat Reverse/Forward Genetics Collection

Location: Cereal Disease Lab

Project Number: 5062-21220-021-27-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Aug 1, 2014
End Date: Jul 31, 2015

The objectives of this project are to: 1. Sequence additional 125 mutant lines using the exon capture platform to expand the sequenced TILLING population. 2. Characterize the chemically induced sequence variation in the exons of this population using our available bioinformatics pipeline. 3. Distribute these sequenced mutants and associated sequence data for further use in analysis of wheat disease resistance and other characters of importance. 4. Screen the lines for changes in disease resistance loci. 5. Add all the new mutations to our current mutant database. 6. Identify candidate disease resistance gene(s) for further molecular analysis. 7. Mutant information and seeds from the additional TILLING lines sequenced as part of this SCA will be publicly available.

Durum cultivar Kronos was mutated with EMS that generates either single nucleotide mutations and or small deletions in the genome. The DNA from mutants will be isolated and exomes for about 82,000 genes will be captured using standard protocol. These exomes will then be sequenced and compared with that of parental lines to determine the mutations in each specific lines. These data will be uploaded into a database for easy search of mutations in specific genes. Additionally, the seeds of these lines will be made available for screening against known rust or Fusarium isolates. Any change from the Kronos parent reaction to a disease would indicate a mutation in a gene involved in host-pathogen interaction. These genes can then be identified by looking at the mutations in that specific line. Further analysis by both groups will allow identification of candidate genes and characterization of the molecular pathways. Thus we can easily go from a gene sequence to a phenotype and/or from a phenotype to a gene sequence utilizing this amazing resource.

Last Modified: 09/21/2017
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