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Construction of a Festuca pratensis BAC library for map-based cloning in Festulolium substitution lines

Donnison, Iain S.; O�Sullivan, Donal M.; Thomas, Ann; Canter, Peter; Moore, Beverley; Armstead, Ian; Thomas, Howard; Edwards, Keith J.; King, Ian P.

Authors

Iain S. Donnison

Donal M. O�Sullivan

Ann Thomas

Peter Canter

Beverley Moore

Ian Armstead

Howard Thomas

Keith J. Edwards



Abstract

Introgression in Festulolium is a potentially powerful tool to isolate genes for a large number of traits which differ between Festuca pratensis Huds. and Lolium perenne L. Not only are hybrids between the two species fertile, but the two genomes can be distinguished by genomic in situ hybridisation and a high frequency of recombination occurs between homoeologous chromosomes and chromosome segments. By a programme of introgression and a series of backcrosses, L. perenne lines have been produced which contain small F. pratensis substitutions. This material is a rich source of polymorphic markers targeted towards any trait carried on the F. pratensis substitution not observed in the L. perenne background. We describe here the construction of an F. pratensis BAC library, which establishes the basis of a map-based cloning strategy in L. perenne. The library contains 49,152 clones, with an average insert size of 112 kbp, providing coverage of 2.5 haploid genome equivalents. We have screened the library for eight amplified fragment length polymorphism (AFLP) derived markers known to be linked to an F. pratensis gene introgressed into L. perenne and conferring a staygreen phenotype as a consequence of a mutation in primary chlorophyll catabolism. While for four of the markers it was possible to identify bacterial artificial chromosome (BAC) clones, the other four AFLPs were too repetitive to enable reliable identification of locus-specific BACs. Moreover, when the four BACs were partially sequenced, no obvious coding regions could be identified. This contrasted to BACs identified using cDNA sequences, when multiple genes were identified on the same BAC.

Citation

Donnison, I. S., O’Sullivan, D. M., Thomas, A., Canter, P., Moore, B., Armstead, I., …King, I. P. (2005). Construction of a Festuca pratensis BAC library for map-based cloning in Festulolium substitution lines. TAG Theoretical and Applied Genetics, 110(5), 846-851. doi:10.1007/s00122-004-1870-y

Journal Article Type Article
Acceptance Date Oct 30, 2004
Online Publication Date Feb 12, 2005
Publication Date 2005-03
Deposit Date Dec 6, 2018
Journal Theoretical and Applied Genetics
Print ISSN 0040-5752
Electronic ISSN 1432-2242
Publisher BMC
Peer Reviewed Peer Reviewed
Volume 110
Issue 5
Pages 846-851
DOI https://doi.org/10.1007/s00122-004-1870-y
Public URL https://nottingham-repository.worktribe.com/output/1379055
Publisher URL https://link.springer.com/article/10.1007%2Fs00122-004-1870-y