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Localization of seed protein genes on flow-sorted field bean chromosomes

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Chromosomes from reconstructed field bean (Vicia faba L.) karyotypes were flow-sorted and the DNA was used for the physical localization of seed storage and nonstorage (USP) protein genes using PCR with sequence specific primers. The data were confirmed and refined by using DNA of microisolated chromosomes of other karyotypes as the target for PCR. The specificity of the PCR products was proved by restrictase digestion into fragments of predicted length or by reamplification using ‘nested’ primers. The genes are located within defined regions of chromosome I (USP=unknown seedprotein genes), II (vicilin genes, legumin B3 genes), III (legumin B4 genes), IV (pseudogenes ψ1) and V (legumin A genes and pseudogenes ψ1). Except for the pseudogene derived from the sequence of legumin B4 gene, all members of each gene family are located in one chromosome region exclusively. This approach proved to be useful for localizing genes that cannot be mapped genetically (due to the lack of allelic variants) and might be applied to integrate physical and genetic maps.

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  1. Arumuganathan K, Slattery JP, Tanksley SD, Earle ED (1991) Preparation and flow cytometric analysis of metaphase chromosomes of tomato.Theor Appl Genet 82: 101–111.

  2. Bäumlein H, Boerjan W, Nagy I,et al. (1991) A novel seed protein gene fromVicia faba is developmentally regulated in transgenic tobacco andArabidopsis plants.Mol Gen Genet 225: 459–467.

  3. Collard JG, de Boer PAJ, Janssen JWG, Schijven JF, de Jong B (1985) Gene mapping by chromosome spot hybridization.Cytometry 6: 179–185.

  4. Cotter F, Nasipuri S, Lam G, Young BD (1989) Gene mapping by enzymatic amplification from flow-sorted chromosomes.Genomics 5: 470–474.

  5. Dellaporta SL, Wood J, Hicks JB (1983) A plant molecular DNA minipreparation. Version II.Plant Mol Biol Rep 1: 19–21.

  6. De Pace C, Delre V, Scarascia Mugnozza GT,et al. (1991) Legumin ofVicia faba major: Accumulation in developing cotyledons, purification, mRNA characterization and chromosomal location of coding genes.Theor Appl Genet 83: 17–23.

  7. Doležel J, Číhalíková J, Lucretti S (1992) A high yield procedure for isolation of metaphase chromosomes from root tips ofVicia faba L.Planta 188: 93–98.

  8. Heim U, Schubert R, Bäumlein H, Wobus U (1989) The legumin gene family: structure and evolutionary implications ofVicia faba B-type genes and pseudogenes.Plant Mol Biol 13: 653–663.

  9. Horstmann C, Schlesier B, Otto A, Kostka S, Müntz K (1993) Polymorphism of legumin subunits from field bean (Vicia faba L. var. minor) and its relation to the corresponding multigene family.Theor Appl Genet 86: in press.

  10. Künzel G, Nicoloff H (1979) Further results on karyotype reconstruction in barley.Biol Zentbl 98: 587–592.

  11. Lucretti S, Doležel J, Schubert I, Fuchs J (1993) Flow karyotyping and sorting ofVicia faba chromosomes.Theor Appl Genet 85: 665–672.

  12. Macas J, Wschke W, Bäumlein H,et al. (1993) Localization of vicilin genes via polymerase chain reaction in microisolated field bean chromosomes.Plant J, in press.

  13. Pich U, Schubert I (1993) Polymorphism of legumin genes in inbred lines ofVicia faba. Biol Zentbl 112: in press

  14. Schlesier B, Bassüner R, Hai NV, Müntz K (1990) The cDNA derived primary structure of two distinct legumin A subunit precursors from field bean (Vicia faba L.).Nucleic Acids Res 18: 7146.

  15. Schubert I, Rieger R (1991) Catalogue of chromosomal and morphological mutants of faba bean in the Gatersleben Collection, 1991.FABIS Newslett 28: 14–22.

  16. Schubert I, Doležel J, Houben A, Scherthan H, Wanner G (1993) Refined examination of plant metaphase chromosome structure at different levels made feasible by new isolation methods.Chromosoma 102: 96–101.

  17. van de Ven M, Powell W, Ramsay G, Waugh R (1990) Restriction fragment length polymorphisms as genetic markers inVicia.Heredity 65: 329–342.

  18. van de Ven WTG, Waugh R, Duncan N, Ramsay G, Dow N, Powell W (1991) Development of a genetic linkage map inVicia faba using molecular and biochemical techniques.Aspects Appl Biol 27: 49–54.

  19. Wang ML, Leitch AR, Schwarzacher T, Heslop-Harrison JS, Moore G (1992) Construction of a chromosome-enrichedHpaII library from flow-sorted wheat chromosomes.Nucleic Acids Res 20: 1897–1901.

  20. Weschke W, Bassüner R, Hai NV, Czihal A, Bäumlein H, Wobus U (1988) The structure of aVicia faba vicilin gene.Biochem Physiol Pflanzen 183: 233–242.

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Correspondence to Ingo Schubert.

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Macas, J., Doležel, J., Lucretti, S. et al. Localization of seed protein genes on flow-sorted field bean chromosomes. Chromosome Res 1, 107–115 (1993). https://doi.org/10.1007/BF00710033

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Key words

  • Vicia faba
  • chromosome flow sorting
  • gene mapping
  • PCR
  • seed protein genes