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Assessing the potential of transcript-based cloning in agriculturally important crop species

Reference	BBS/E/J/0000A256
Principal Investigator / Supervisor	Dr Lesley Boyd
Co-Investigators / Co-Supervisors	Dr Margaret Boulton
Institution	John Innes Centre
Department	John Innes Centre Department
Funding type	Research
Value (£)	7,477
Status	Completed
Type	Institute Project
Start date	01/10/2005
End date	31/10/2006
Duration	13 months

Abstract

Transcript-based cloning (TBC), utilising transcript-defective mutants, has proven a viable approach to isolate genes from the diploid genomes of Medicago truncatula and barley. To make this method of gene identification of greater utility to the agricultural industry we have to address the key question pertaining to gene cloning in crop species: Is TBC a viable approach to gene isolation in complex genomes such as that of our major cereal crop, hexaploid bread wheat. This proposal will address this questions using the following materials and approaches. Yr5 confers resistance in wheat to yellow rust. Three mutant alleles of Yr5 have been generated by fast neutrons, a mutagen known to cause deletions. A genetic marker that co-segregates with Yr5 indicates that all three alleles are indeed deletions. Using the wheat Affymetrix microarray we will screen for genes down-regulated in the three Yr5 mutant alleles compared with wild type wheat. The gene(s) down-regulated in all the Yr5 mutant alleles will be further analysed. We have good evidence from comparative mapping to rice that Yr5 encodes for a NBS-LRR type R-gene, so this prior knowledge will help in the identification of Yr5 candidates. We will verify Yr5 candidates by screening for co-segregation with the Yr5 phenotype in pre-existing mapping populations. Successful identification of Yr5 will indicate that transcript-based cloning is viable in wheat. Considering that wheat has one of the most complex genomes of all crop species we can infer that this technology is most likely viable in all crop plants.

Summary

unavailable

Committee	Closed Committee - Plant & Microbial Sciences (PMS)
Research Topics	X – not assigned to a current Research Topic
Research Priority	X – Research Priority information not available
Research Initiative	X - not in an Initiative
Funding Scheme	X – not Funded via a specific Funding Scheme

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