title: Comparative genomics of Lupinus angustifolius gene-rich regions: BAC library exploration, genetic mapping and cytogenetics.
abstract: BACKGROUND
  The narrow-leafed lupin, Lupinus angustifolius L., is a grain legume species with a relatively compact genome. The species has 2n = 40 chromosomes and its genome size is 960 Mbp/1C. During the last decade, L. angustifolius genomic studies have achieved several milestones, such as molecular-marker development, linkage maps, and bacterial artificial chromosome (BAC) libraries. Here, these resources were integratively used to identify and sequence two gene-rich regions (GRRs) of the genome.
  
  
  RESULTS
  The genome was screened with a probe representing the sequence of a microsatellite fragment length polymorphism (MFLP) marker linked to Phomopsis stem blight resistance. BAC clones selected by hybridization were subjected to restriction fingerprinting and contig assembly, and 232 BAC-ends were sequenced and annotated. BAC fluorescence in situ hybridization (BAC-FISH) identified eight single-locus clones. Based on physical mapping, cytogenetic localization, and BAC-end annotation, five clones were chosen for sequencing. Within the sequences of clones that hybridized in FISH to a single-locus, two large GRRs were identified. The GRRs showed strong and conserved synteny to Glycine max duplicated genome regions, illustrated by both identical gene order and parallel orientation. In contrast, in the clones with dispersed FISH signals, more than one-third of sequences were transposable elements. Sequenced, single-locus clones were used to develop 12 genetic markers, increasing the number of L. angustifolius chromosomes linked to appropriate linkage groups by five pairs.
  
  
  CONCLUSIONS
  In general, probes originating from MFLP sequences can assist genome screening and gene discovery. However, such probes are not useful for positional cloning, because they tend to hybridize to numerous loci. GRRs identified in L. angustifolius contained a low number of interspersed repeats and had a high level of synteny to the genome of the model legume G. max. Our results showed that not only was the gene nucleotide sequence conserved between soybean and lupin GRRs, but the order and orientation of particular genes in syntenic blocks was homologous, as well. These findings will be valuable to the forthcoming sequencing of the lupin genome.
authors: Institute of Plant Genetics, Polish Academy of Sciences, Strzeszyńska 34, 60-479 Poznań, Poland. mksi@igr.poznan.pl
authors: Michał
authors: M
authors: Książkiewicz
authors: Katarzyna
authors: K
authors: Wyrwa
authors: Anna
authors: A
authors: Szczepaniak
authors: Sandra
authors: S
authors: Rychel
authors: Karolina
authors: K
authors: Majcherkiewicz
authors: Łucja
authors: Ł
authors: Przysiecka
authors: Wojciech
authors: W
authors: Karlowski
authors: Bogdan
authors: B
authors: Wolko
authors: Barbara
authors: B
authors: Naganowska
mesh_terms: D002874:Chromosome Mapping; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics*; D020451:Contig Mapping; D003582:Cytogenetics; D004251:DNA Transposable Elements / Q000235:genetics; D015723:Gene Library; D008040:Genetic Linkage; D005819:Genetic Markers / Q000235:genetics; D018745:Genome, Plant; D017404:In Situ Hybridization, Fluorescence; D031297:Lupinus / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D017422:Sequence Analysis, DNA
chemical_list: D004251:DNA Transposable Elements; D005819:Genetic Markers
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