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Title  Genomics-assisted dissection of barley morphology and development
   
Acronym BARCODE
   
Duration 1 November 2007 - 1 November 2010
   
Project leader Robbie Waugh, Scottisch Crop Research Institute, United Kingdom
   

Other project participants

Niels Stein, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Germany
Michele Morgante, University of Udine, Italy
  
Funding The German Research Foundation (DFG), Germany  
  Ministry of Education, University and Research (MIUR), Italy
  Biotechnological and Biological Sciences Research Council (BBSRC), United Kingdom
   
Total granted budget  € 1,532,940
   
Abstract  

We will use Illumina Oligo Pool Assay (OPA) technology to genotype 480 Bowman backcross derived nearly isogenic mutant lines at 3072 mapped genetic loci (6144 alleles). We will use simple pair-wise comparison with Bowman, their recurrent parent, to define the boundaries of the segments containing mutated alleles and compile a detailed comparative putative gene content map for each locus based on the barley, rice and emerging Brachypodium and maize genome sequences. While OPA genotyping will narrow the location of each mutant locus to a small genetic interval, identifying the mutated gene from all potential candidates will likely still require a forward genetics approach. To facilitate this, and all similar studies in the future, we propose to contribute to an established international strategy for the staged development of a genetically anchored physical map of the barley genome by end sequencing BACs that are currently undergoing High Information Content Fingerprint analysis and Overgo hybridisation to >12,000 unigenes. This will provide up to 400 Mbp of barley genomic DNA sequence information and will assist in anchoring the physical map to the barley gene map (~5-6000 mapped genes) and rice genome sequence. To demonstrate the utility of the developed information, resources and approach, each partner lab will identify a minimum BAC tiling path spanning a morphological /developmental mutant locus, identify candidate genes and initiate their functional characterisation by a combination of comparative allele sequencing (WT vs. Mutant) and the use of functional genomics tools available to the group (TILLING, transgenics, VIGS). All information will be made publicly available to the community for broader exploitation through existing web-based informatics resources.

   
Progress
  • Genotyping the Bowman NILS has been completed (3060 SNPs, 979 lines) and data is being drafted for publication
  • Over 300,000 Paired BAC End Sequences have been derived and are being incorporated into the barley physical map being constructed out with this project.
  • 27 F2 populations of >10,000 individuals have been developed for a range of mutant phenotypes and distributed to IPK (9), Udine (9) and SCRI (9).
    Fine mapping of three chosen morphological mutant loci is underway using the homozygous recessive individuals of large F2 populations
 
Figure 1 Breeding Figure 2 In vitro
  Figure 3 Backcross populations  Figure 4 Transient gene silencing  
Project website

http://pgrc.ipk-gatersleben.de/barleynet/projects_barcode.php