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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Allouis S, Moore G, Bellec A, Sharp R, Faivre Rampant P, Mortimer K, Pateyron S, Foote TN, Griffiths S, Caboche M, Chalhoub B (2003) Construction and characterisation of a hexaploid wheat (Triticum aestivum L.) BAC library from the reference germplasm ‘Chinese Spring’. Cereal Res Commun 31:331–338
Armstead IP, Bollard A, Morgan WG, Harper JA, King IP, Jones RN, Forster JW, Hayward MD, Thomas HM (2001) Genetic and physical analysis of a single Festuca pratensis chromosome segment substitution in Lolium perenne. Chromosoma 110:52–57
Armstead IP, Turner LB, Farrell M, Skot L, Gomez P, Montoya T, Donnison IS, King IP, Humphreys MO (2004) Synteny between a major heading-date QTL in perennial ryegrass (Lolium perenne L.) and the Hd3 heading-date locus in rice. Theor Appl Genet 108:822–828
Bennett MD, Smith JB, Heslop-Harrison JS (1982) Nuclear DNA amounts in angiosperms. Proc R Soc London Ser Biol 216:179–199
Cenci A, Chantret N, Kong X, Gu Y, Anderson OD, Fahima T, Distelfeld A, Dubcovsky J (2003) Construction and characterization of a half million clone BAC library of durum wheat (Triticum turgidum ssp durum). Theor Appl Genet 107:931–939
Chalhoub B, Belcram H, Caboche M (2004) Efficient cloning of plant genomes into bacterial artificial chromosome (BAC) libraries with larger and more uniform insert size. Plant Biotechnol J 2:181–188
Clarke L, Carbon J (1976) A colony bank containing synthetic Col El hybrid plasmids representative of the entire E. coli genome. Cell 9:91–99
Humphreys MW, Humphreys J, Donnison IS, King IP, Thomas HM, Guesquiere M, Durand J-E, Rognli O-A, Zwierzykowski Z, Rapacz M (2004) Molecular breeding and functional genomics for tolerance to abiotic stress. In: Hopkins A, Wang Z-Y, Mian R, Sledge M, Barker RE (eds) Molecular breeding of forage and turf. Developments in plant breeding, vol 11. Kluwer, Dordrecht, pp 61–80
King IP, Morgan WG, Armstead IP, Harper JA, Hayward MD, Bollard A, Nash JV, Forster JW, Thomas HM (1998a) Introgression mapping in the grasses I. Introgression of Festuca pratensis chromosomes and chromosome segments into Lolium perenne. Heredity 81:462–467
King IP, Morgan WG, Harper JA, Thomas HM (1998b). Introgression mapping in the grasses II. Meiotic analysis of the Lolium perenne/Festuca pratensis triploid hybrid. Heredity 82:107–112
King J, Roberts LA, Kearsey MJ, Thomas HM, Jones RN, Huang L, Armstead IP, Morgan WG, King IP (2002a) A demonstration of a 1:1 correspondence between chiasma frequency and recombination using a Lolium perenne/Festuca pratensis substitution line. Genetics 161:315–324
King J, Armstead IP, Donnison IS, Thomas HM, Jones RN, Kearsey MJ, Roberts LA, Thomas A, Morgan WG, King IP (2002b) Physical and genetic mapping in the grasses Lolium perenne and Festuca pratensis. Genetics 161:315–324
O’Sullivan DM, Ripoll PJ, Rodgers M, Edwards KJ (2001) A maize bacterial artificial chromosome (BAC) library from the European flint inbred line F2. Theor Appl Genet 103:425–432
Roca M, James CL, Pruzinska A, Hortensteiner S, Thomas H, Ougham H (2004) Analysis of the chlorophyll catabolism pathway in leaves of an introgression senescence mutant of Lolium temulentum. Phytochemistry 65:1231–1238
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Press, Cold Spring Harbor
Shizuya H, Birren B, Kim UJ, Mancino V, Slepak T, Tachiiri Y, Simon M (1992) Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an f-factor-based vector. Proc Natl Acad Sci USA 89:8794–8797
Thomas H, Evans C, Thomas HM, Humphreys MW, Morgan G, Hauck B, Donnison IS (1997) Introgression, tagging and expression of a leaf senescence gene in Festulolium. New Phytol 137:29–34
Tomkins JP, Davis G, Main D, Yim Y, Duru N, Musket T, Goicoechea JL, Frisch DA, Coe EH, Wing RA (2002) Construction and characterization of a deep-coverage bacterial artificial chromosome library for maize. Crop Sci 42:928–933
Vicentini F, Hortensteiner S, Schellenberg M, Thomas H, Matile P (1995) Chlorophyll breakdown in senescent leaves: identification of the biochemical lesion in a stay-green genotype of Festuca pratensis. New Phytol 129:247–252
Woo SS, Jiang JM, Gill BS, Paterson AH, Wing RA (1994) Construction and characterization of a bacterial artificial chromosome library of Sorghum bicolor. Nucleic Acids Res 22:4922–4931
Yu Y, Tomkins JP, Waugh R, Frisch DA, Kudrna D, Kleinhofs A, Brueggeman RS, Muehlbauer GJ, Wise RP, Wing RA (2000) A bacterial artificial chromosome library for barley (Hordeum vulgare L.) and the identification of clones containing putative resistance genes. Theor Appl Genet 101:1093–1099
Zhang HB, Zhao XP, Ding XL, Paterson AH, Wing RA (1995) Preparation of megabase-size DNA from plant nuclei. Plant J 7:175–184
Zhang HB, Choi SD, Woo SS, Li ZK, Wing RA (1996) Construction and characterization of two rice bacterial artificial chromosome libraries from the parents of a permanent recombinant inbred mapping population. Mol Breed 2:11–24
Acknowledgements
The Institute of Grassland and Environmental Research (IGER) is sponsored by the UK Biotechnology and Biological Sciences Research Council of the United Kingdom. Iain Donnison is grateful to Pierre Jean Ripoll for helpful advice during BAC library construction. The authors also wish to thank Simon Bright, Mary Knight and Helene Lacroix at Syngenta for the use of their BioPick (BioRobotics) to pick some of the BAC clones. Requests to screen the library should be addressed to Iain Donnison (e-mail: iain.donnison@bbsrc.ac.uk).
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by J.W. Snape
Rights and permissions
About this article
Cite this article
Donnison, I.S., O’Sullivan, D.M., Thomas, A. et al. Construction of a Festuca pratensis BAC library for map-based cloning in Festulolium substitution lines. Theor Appl Genet 110, 846–851 (2005). https://doi.org/10.1007/s00122-004-1870-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00122-004-1870-y